Phytopathogenic organisms and mycotoxigenic fungi: Why do we control one and neglect the other? A biological control perspective in Malaysia

In this review, we present the current information on development and applications of biological control against phytopathogenic organisms as well as mycotoxigenic fungi in Malaysia as part of the integrated pest management (IPM) programs in a collective effort to achieve food security. Although the biological control of phytopathogenic organisms of economically important crops is well established and widely practiced in Malaysia with considerable success, the same cannot be said for mycotoxigenic fungi. This is surprising because the year round hot and humid Malaysian tropical climate is very conducive for the colonization of mycotoxigenic fungi and the potential contamination with mycotoxins. This suggests that less focus has been made on the control of mycotoxigenic species in the genera Aspergillus, Fusarium, and Penicillium in Malaysia, despite the food security and health implications of exposure to the mycotoxins produced by these species. At present, there is limited research in Malaysia related to biological control of the key mycotoxins, especially aflatoxins, Fusarium‐related mycotoxins, and ochratoxin A, in key food and feed chains. The expected threats of climate change, its impacts on both plant physiology and the proliferation of mycotoxigenic fungi, and the contamination of food and feed commodities with mycotoxins, including the discovery of masked mycotoxins, will pose significant new global challenges that will impact on mycotoxin management strategies in food and feed crops worldwide. Future research, especially in Malaysia, should urgently focus on these challenges to develop IPM strategies that include biological control for minimizing mycotoxins in economically important food and feed chains for the benefit of ensuring food safety and food security under climate change scenarios.     

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.     

天然植物成分防控农产品中真菌毒素的研究进展

真菌毒素是真菌生长过程中产生的次生代谢产物,其对农产品的污染直接威胁人类和动物的生命健康。真菌毒素的预防和脱除是实现食品和饲料工业高质量发展亟待解决的关键问题之一。目前研究者采用了多种策略来防控真菌毒素污染避免健康问题和经济损失,包括抑制真菌生长及真菌毒素生成、去除和降解污染农产品中的真菌毒素、降低真菌毒素生物活性等。利用天然植物成分（Natural plant compounds,NPC）防控真菌毒素污染表现出稳定性强、安全性好和抑制效率高等优势,业已成为研究新趋势。本文综述了近年来NPC防控农产品中真菌毒素污染的不同策略,讨论了相应的作用机制,分析了现阶段采用NPC防控真菌毒素的优势和不足,并展望了在食品工业的应用前景,为开发新的真菌毒素防控试剂提供科学参考。     

Predicting mycotoxins in foods: A review

The need to ensure the microbiological quality and safety of food products has stimulated interest in the use of mathematical models for quantifying and predicting microbial behaviour. For 20 years, predictive microbiology has been developed for predicting the occurrence of food-borne pathogens, although these tools are dedicated to bacteria. Recently, the situation has changed and a growing number of studies are available in the literature dealing with the predictive modelling approach of fungi. To our knowledge the present one is the first review focussed on predictive mycology and food safety, including mycotoxins; existing kinetic and probability models applied to mycotoxigenic fungi germination and growth, and mycotoxin production are reviewed.     

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 and preventive strategies to control mycotoxins in cereal‐based food

Mycotoxins contamination in cereal‐based food is ubiquitous according to systematic review of the scientific documentation of worldwide mycotoxin contamination in cereal and their products between 2008 and 2018, thus representing food safety issue especially in developing tropical countries. Food processing plays a vital role to prevent mycotoxin contamination in food. Therefore, it is with great urgency to develop strategies to inhibit fungi growth and mycotoxin production during food processing. This review begins by discussing physicochemical properties of five most common mycotoxins (aflatoxins, fumonisins, ochratoxins, deoxynivalenol, and zearalenone) found in cereal grains, regulation for mycotoxins in food, and their potential negative impact on human health. The fate of mycotoxins during major cereal‐based food processing including milling, breadmaking, extrusion, malting, and brewing was then summarized. In the end, traditional mitigation strategies including physical and chemical and potential application of biocontrol agent and essential oil nanoemulsions that can be applied during food processing were discussed. It indicated that no single method is currently available to completely prevent mycotoxin contamination in cereal foods.     

Mycotoxins in dairy products

Certain fungi produce chemical substances that cause toxic symptoms when food containing them is ingested by man or animals. These compounds are referred to as mycotoxins. Mycotoxins may contaminate dairy products by moulds growing on them, or by the carry-over of mycotoxins occurring in animal feedstuffs ingested by dairy cattle. An example of the first mentioned category is sterigmatocystin, a carcinogenic mycotoxin sometimes occurring on hard cheese. An example of the second category is aflatoxin M₁, a compound strongly suspected to be carcinogenic, which often occurs in milk. Due to the fact that processing of milk does not decrease the aflatoxin M₁ content, aflatoxin M₁ occurs in various dairy products. Sensitive methods of analysis for the determination of mycotoxins in dairy products have been developed in the last 10 years, most of them are based on TLC- or HPLC-separation procedures, followed by fluorimetric measurement.The most fundamental way to tackle the problem of mycotoxin contamination of dairy products is to prevent fungal growth on the dairy products or, in the case of carry-over of mycotoxins, in the crop before, during and after harvest. If measures to prevent fungal growth and mycotoxins production are not taken or fail, one can sometimes resort to physical or chemical methods to eliminate mycotoxins.     

Novel non-thermal food processing techniques and their mechanism of action in mycotoxins decontamination of foods

This review describes the major food and feed contaminating mycotoxins and provides a thorough insight about non-thermal food processing techniques and their mycotoxin detoxification mechanisms. Cold plasma, pulsed light, pulsed electric field, high pressure processing, and electron beam irradiation are among the techniques discussed. Mycotoxins decontamination is usually achieved through the release of reactive species and inactivation of toxin-producing microorganisms through alteration of cell membrane integrity and genetic makeup. Destruction of the molecular structure of mycotoxins responsible for toxicity also occurs during these processes. These non-thermal methods are effective in decontaminating mycotoxins with varying degrees of efficiency, and some of the methods do complete decontamination of mycotoxins with minimal processing. Despite their promising efficacy in decontaminating mycotoxins, the feasibility of most of these methods requires scale-up with future potential for commercialization and acceptance. Efforts should be made to increase the scalability and adoption of the technologies, especially in low-income countries where mycotoxin contamination is prevalent.     

Use of essential oils and phytochemicals against the mycotoxins producing fungi for shelf-life enhancement and food preservation

Mycotoxin-producing fungi are a significant source of crop and food contamination, posing a significant threat to global food safety and security. Essential oils, plant extracts and phytochemicals have emerged as green preservatives to extend the shelf-life of foods due to their unique antimicrobial properties. Unlike conventional synthetic preservatives, they are a sustainable and safe way to preserve food with no or little harmful effects on the environment. Use of nanoformulations containing essential oils and phytochemicals offer enormous potential as a mitigation strategy to lower mycotoxin contamination incidences in food and crop with enhanced release behaviour to efficiently transport them to the target location for a rapid reaction without much impact from environmental variables. Hence, this review overviews various essential oils and phytochemicals utilized through nanoformulations to control the mycotoxigenic fungi, probable mechanism of actions involved as well as emerging mycotoxins and associated safety concerns to ensure food sustainability.     

Molecular biodiversity of mycotoxigenic fungi that threaten food safety

Fungal biodiversity is one of the most important contributors to the occurrence and severity of mycotoxin contamination of crop plants. Phenotypic and metabolic plasticity has enabled mycotoxigenic fungi to colonize a broad range of agriculturally important crops and to adapt to a range of environmental conditions. New mycotoxin-commodity combinations provide evidence for the ability of fungi to adapt to changing conditions and the emergence of genotypes that confer enhanced aggressiveness toward plants and/or altered mycotoxin production profiles. Perhaps the most important contributor to qualitative differences in mycotoxin production among fungi is variation in mycotoxin biosynthetic genes. Molecular genetic and biochemical analyses of toxigenic fungi have elucidated specific differences in biosynthetic genes that are responsible for intra- and inter-specific differences in mycotoxin production. For Aspergillus and Fusarium, the mycotoxigenic genera of greatest concern, variation in biosynthetic genes responsible for production of individual families of mycotoxins appears to be the result of evolutionary adaptation. Examples of such variation have been reported for: a) aflatoxin biosynthetic genes in Aspergillus flavus and Aspergillus parasiticus; b) trichothecene biosynthetic genes within and among Fusarium species; and c) fumonisin biosynthetic genes in Aspergillus and Fusarium species. Understanding the variation in these biosynthetic genes and the basis for variation in mycotoxin production is important for accurate assessment of the risks that fungi pose to food safety and for prevention of mycotoxin contamination of crops in the field and in storage.     

Impact of nonthermal food processing techniques on mycotoxins and their producing fungi

Mycotoxins are a significant threat to food safety and quality. Over the years, mycotoxins have been detected in almost all food and feed crops without any regional barrier. Conventional techniques for decontamination of mycotoxin involve physical, chemical, and biological methods, but these technologies often impact the quality of food in terms of changes in nutritional and sensory attributes. We examined the effects of nonthermal techniques on mycotoxins and their producing fungi to remove or reduce mycotoxin levels in food products without compromising food quality. Nonthermal technologies employ different lethal agents (including ozone, cold plasma, light, pressure, radiation, ultrasound, electric field, and magnetic field) to degrade mycotoxins while minimising product thermal exposure. However, the degradation pathway and toxicology of treated products need further research for a better understanding. With such food process development and optimisation efforts, food processors can employ various nonthermal technologies as tools for delivering consumer-desired mycotoxin-free food products with intact nutritional and sensory quality.     

Recent developments and applications of hyperspectral imaging for rapid detection of mycotoxins and mycotoxigenic fungi in food products

Mycotoxins are the foremost naturally occurring contaminants of food products such as corn, peanuts, tree nuts, and wheat. As the secondary metabolites, mycotoxins are mainly synthesized by many species of the genera Aspergillus, Fusarium and Penicillium, and are considered highly toxic and carcinogenic to humans and animals. Most mycotoxins are detected and quantified by analytical chemistry-based methods. While mycotoxigenic fungi are usually identified and quantified by biological methods. However, these methods are time-consuming, laborious, costly, and inconsistent because of the variability of the grain-sampling process. It is desirable to develop rapid, non-destructive and efficient methods that objectively measure and evaluate mycotoxins and mycotoxigenic fungi in food. In recent years, some spectroscopy-based technologies such as hyperspectral imaging (HSI), Raman spectroscopy, and Fourier transform infrared spectroscopy have been extensively investigated for their potential use as tools for the detection, classification, and sorting of mycotoxins and toxigenic fungal contaminants in food. HSI integrates both spatial and spectral information for every pixel in an image, making it suitable for rapid detection of large quantities of samples and more heterogeneous samples and for in-line sorting in the food industry. In order to track the latest research developments in HSI, this paper gives a brief overview of the theories and fundamentals behind the technology and discusses its applications in the field of rapid detection and sorting of mycotoxins and toxigenic fungi in food products. Additionally, advantages and disadvantages of HSI are compared, and its potential use in commercial applications is reported.     

Lactic Acid Bacteria as Antifungal and Anti‐Mycotoxigenic Agents: A Comprehensive Review

Fungal contamination of food and animal feed, especially by mycotoxigenic fungi, is not only a global food quality concern for food manufacturers, but it also poses serious health concerns because of the production of a variety of mycotoxins, some of which present considerable food safety challenges. In today's mega‐scale food and feed productions, which involve a number of processing steps and the use of a variety of ingredients, fungal contamination is regarded as unavoidable, even good manufacturing practices are followed. Chemical preservatives, to some extent, are successful in retarding microbial growth and achieving considerably longer shelf‐life. However, the increasing demand for clean label products requires manufacturers to find natural alternatives to replace chemically derived ingredients to guarantee the clean label. Lactic acid bacteria (LAB), with the status generally recognized as safe (GRAS), are apprehended as an apt choice to be used as natural preservatives in food and animal feed to control fungal growth and subsequent mycotoxin production. LAB species produce a vast spectrum of antifungal metabolites to inhibit fungal growth; and also have the capacity to adsorb, degrade, or detoxify fungal mycotoxins including ochratoxins, aflatoxins, and Fusarium toxins. The potential of many LAB species to circumvent spoilage associated with fungi has been exploited in a variety of human food and animal feed stuff. This review provides the most recent updates on the ability of LAB to serve as antifungal and anti‐mycotoxigenic agents. In addition, some recent trends of the use of LAB as biopreservative agents against fungal growth and mycotoxin production are highlighted.     

Advanced synchrotron-based and globar-sourced molecular (micro) spectroscopy contributions to advances in food and feed research on molecular structure,mycotoxin determination,and molecular nutrition

ABSTRACT

Mycotoxin contamination has been a worldwide problem for food and feeds production for a long time. There is an obviously increased focus of the food and feed industry toward the reduction of mycotoxin concentration in both the raw materials and finished products. Therefore, both effective qualitative and quantitative techniques for the determination of mycotoxins are required to minimize their harmful effects. Conventional wet chemical methods usually are time-consuming, expensive, and rely on complex extraction and cleanup pretreatments. Synchrotron-based and globar-based molecular spectroscopy have shown great potential to be developed as rapid and nondestructive tools for the determination of molecular structure, molecular nutrition and mycotoxins in feed and food. This article reviews the common types of mycotoxins in feed and food, their toxicity, as well as the conventional detection methods. The principle of advanced molecular spectroscopy techniques and their application prospects for mycotoxin detection are discussed. Recent progress in food and feed research with molecular spectroscopy techniques is highlighted. This review provides a potential and insight into how to determine the structure and mycotoxins of feed and food on a molecular basis with advanced Synchrotron-based and globar-based molecular (micro) spectroscopy.     

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.     

Post-harvest control strategies: minimizing mycotoxins in the food chain

Contamination of cereal commodities by moulds and mycotoxins results in dry matter, quality, and nutritional losses and represents a significant hazard to the food chain. Most grain is harvested, dried and then stored on farm or in silos for medium/long term storage. Cereal quality is influenced by a range of interacting abiotic and biotic factors. In the so-called stored grain ecosystem, factors include grain and contaminant mould respiration, insect pests, rodents and the key environmental factors of temperature, water availability and intergranular gas composition, and preservatives which are added to conserve moist grain for animal feed. Thus knowledge of the key critical control points during harvesting, drying and storage stages in the cereal production chain are essential in developing effective prevention strategies post-harvest. Studies show that very small amounts of dry matter loss due to mould activity can be tolerated. With <0.5% dry matter loss visible moulding, mycotoxin contamination and downgrading of lots can occur. The key mycotoxigenic moulds in partially dried grain are Penicillium verrucosum (ochratoxin) in damp cool climates of Northern Europe, and Aspergillus flavus (aflatoxins), A. ochraceus (ochratoxin) and some Fusarium species (fumonisins, trichothecenes) on temperate and tropical cereals. Studies on the ecology of these species has resulted in modelling of germination, growth and mycotoxin minima and prediction of fungal contamination levels which may lead to mycotoxin contamination above the tolerable legislative limits (e.g. for ochratoxin). The effect of modified atmospheres and fumigation with sulphur dioxide and ammonia have been attempted to try and control mould spoilage in storage. Elevated CO2 of >75% are required to ensure that growth of mycotoxigenic moulds does not occur in partially dried grain. Sometimes, preservatives based on aliphatic acids have been used to prevent spoilage and mycotoxin contamination of stored commodities, especially feed. These are predominantly fungistats and attempts have been made to use alternatives such as essential oils and anti-oxidants to prevent growth and mycotoxin accumulation in partially dried grain. Interactions between spoilage and mycotoxigenic fungi and insect pests inevitably occurs in stored grain ecosystems and this can further influence contamination with mycotoxins. Effective post-harvest management of stored commodities requires clear monitoring criteria and effective implementation in relation to abiotic and biotic factors, hygiene and monitoring to ensure that mycotoxin contamination is minimised and that stored grain can proceed through the food chain for processing.     

How will climate change affect mycotoxins in food?

This invited review and opinion piece, assesses the impact of climate change on mycotoxins in food: only one paper and an abstract referred directly from a substantial literature search and then only in relation to Europe. Climate change is an accepted probability by most scientists. Favourable temperature and water activity are crucial for mycotoxigenic fungi and mycotoxin production. Fungal diseases of crops provide relevant information for pre-harvest mycotoxin contamination. However, the mycotoxin issue also involves post-harvest scenarios. There are no data on how mycotoxins affect competing organisms in crop ecosystems. In general, if the temperature increases in cool or temperate climates, the relevant countries may become more liable to aflatoxins. Tropical countries may become too inhospitable for conventional fungal growth and mycotoxin production. Could this lead to the extinction of thermotolerant Aspergillus flavus? Currently cold regions may become liable to temperate problems concerning ochratoxin A, patulin and Fusarium toxins (e.g. deoxynivalenol). Regions which can afford to control the environment of storage facilities may be able to avoid post-harvest problems but at high additional cost. There appears to be a lack of awareness of the issue in some non-European countries. The era will provide numerous challenges for mycotoxicologists.     

A review on novel non-thermal food processing techniques for mycotoxin reduction

The overarching challenges of mycotoxin contamination in food necessitate the development of strategies to be implemented to combat their effects thereof. Common processing techniques have been utilised but do not necessarily meet the desired efficacy. This review appraises studies on novel non-thermal food processing techniques, particularly high pressure processing, pulsed electric filed, cold plasma and ultrasound processing for the decontamination of mycotoxins in food. Although available studies on these techniques have suggested a reduction of mycotoxins and in some instances, complete decontamination of mycotoxins was also reported. The mechanisms by which reduction/elimination occurs include through decomposition of toxins after collision with ions/electrons leading to cleavage of bonds, structural degradation of the mycotoxins structure and cleavage of functional groups. Additional studies into the toxicity of degraded products and the composition of the food products are still required to ensure a more widespread adoption of these techniques to enhance food safety.     

Integrated management of the risks of stored grain spoilage by seedborne fungi and contamination by storage mould mycotoxins – An update

Fungal spoilage of stored grains may occur when activity of water (aw) in cereal grain exceeds a critical limit enabling mould growth. Because it is not feasible to maintain all parts of large grain bulks below this critical moisture limit during prolonged storage time, an infection by seed-borne fungi is not rare in cereal grain stored under humid temperate or hot climates, inducing irreversible qualitative losses. Additionally, some fungal species produce harmful mycotoxins. The most harmful toxigenic species belong to the group of xerophilic species (genera Aspergillus and Penicillium). Because mycotoxin contamination of cereal grain is a worldwide issue for public health and a permanent concern for cereal-food industries facing the challenge of a permanent monitoring mycotoxin content in their primary matters, tolerable levels of mycotoxins are severely regulated worldwide. Mycotoxin-producing species growth is closely dependent of grain moisture levels enabling biological activity in grain ecosystem. Consequently, mould growth in stored grain bulks can be anticipated through early detection of grain and mould respiration. The prevention of mycotoxigenic fungi spoilage of stored grain can be managed by a preventive strategy. The main objective of the review was to describe the different methods, material and practices combined in such an integrated preventive approach. Some solutions potentially acceptable for the decontamination of moderately contaminated grain are also discussed.Integrated management of mould spoilage risks in stored grain is based on five pillars: i/Prevention of mould development by keeping grain moisture below the critical limit of fungal growth; ii/Accurate monitoring of grain aw and temperature changes during the storage period, associated to the monitoring of early indicators of respiration activity of storage fungi; iii/Reduction of grain bulk moistening trends by physical intervention means; iv/Use of physical treatments (ozone, grain peeling or abrasion) to limit mycotoxin contamination transfer to processed cereal products; v/Possible use of bio-competitive strains of fungi or bacteria to prevent the development of mycotoxigenic fungi in grain bulks. The future research needs on this topic are also evocated.     

Review of mycotoxin reduction in food and feed: from prevention in the field to detoxification by adsorption or transformation

Mycotoxins are secondary metabolites present worldwide in agricultural commodities and produced by ?lamentous fungi that cause a toxic response (mycotoxicosis) when ingested by animals. Prevention of mycotoxicoses includes pre- and post-harvest strategies. The best way to reduce the mycotoxin content in food and feed is the prevention of mycotoxin formation in the ?eld, but this is often not sufficient, so other methods are needed. To decontaminate and/or detoxify mycotoxin-contaminated food and feed, the most prevalent approach in the feed industry is the inclusion of sorbent materials in the feed thus obtaining more or less selective removal of toxins by adsorption during passage through the gastrointestinal tract. Another reliable approach is to add enzymes or microorganisms capable of detoxifying some mycotoxins. Through a comprehensive review of published reports on the strategies for mycotoxin removal, this present work aims to update our understanding of mycotoxin removal. It provides an insight into the detoxification of mycotoxin present in food and feed. In the future, more emphasis needs to be placed on adsorption of mycotoxins in the gastrointestinal tract. Concerning the enzymatic transformation of mycotoxins, further efforts are required in understanding detoxification reactions, the toxicity of transformation products and in the characterization of enzymes responsible for transformations.