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.     

Limiting mycotoxins in stored wheat

The quality of harvested wheat grain can deteriorate markedly during the post-harvest management stages. Biotic factors, such as grain type and ripeness, coupled with the prevailing abiotic factors, such as water content and temperature, and also preservative concentration will influence the safe storage life and the level of contamination with mycotoxins. These mycotoxins include deoxynivalenol (DON) produced pre-harvest and zearalenone (ZEA) produced post-harvest by Fusarium graminearum and Fusarium poae, respectively, ochratoxin (OTA) produced by Penicillium verrucosum post-harvest in cool damp northern European climates, and perhaps T-2 and HT-2 toxins produced by Fusarium langsethiae. This review presents recent data on the relationship between dry matter losses caused by F. graminearum under different environmental regimes (water activities, temperatures) and the level of contamination with DON. This is important as poor post-harvest drying and storage management may exacerbate DON contamination already present pre-harvest. It is thus critical to relate the environmental factors in stored wheat grain during storage, especially of intergranular relative humidity (RH) and temperature, to safe storage periods without spoilage or risk from increased DON contamination. The growth/no growth and DON/no DON (F. graminearum) and OTA/no toxin production (P. verrucosum) have been used to build a model with a simple interface to link temperature and RH values to the potential risk level which may allow growth or toxin production. This paper also considers the use of modified atmospheres, preservatives and biocontrol to minimise DON and OTA in moist wheat grain. These approaches together with clear monitoring criteria and hygiene could contribute to better post-harvest management of stored temperate cereals and ensure that mycotoxin contamination is minimised during this key phase in the food/feed chain.     

Novel approaches for chemical and microbiological shelf life extension of cereal crops

Abstract

Economic losses due to post-harvest fungal spoilage and mycotoxin contamination of cereal crops is a frequently encountered issue. Typically, chemical preservatives are used to reduce the initial microbial load and the environmental conditions during storage are controlled to prevent microbial growth. However, in recent years the consumers’ desire for more naturally produced foods containing less chemical preservatives has grown increasingly stronger. This article reviews the latest advances in terms of novel approaches for chemical decontamination, namely application cold atmospheric pressure plasma and electrolyzed water, and their suitability for preservation of stored cereal crops. In addition, the alternative use of bio-preservatives, such as starter cultures or purified antimicrobial compounds, to prevent the growth of spoilage organisms or remove in-field accumulated mycotoxins is evaluated. All treatments assessed here show potential for inhibition of microbial spoilage. However, each method encounters draw-backs, making industrial application difficult. Even under optimized processing conditions, it is unlikely that one single treatment can reduce the natural microbial load sufficiently. It is evident that future research needs to examine the combined application of several treatments to exploit their synergistic properties. This would enable sufficient reduction in the microbial load and ensure microbiological safety of cereal crops during long-term storage.     

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.     

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.     

Effects of fungal deterioration on grain: nutritional value, toxicity, germination

Moulds or fungi that grow in grains and seeds during storage and transport cause germination decrease, visible mouldiness, discoloration, musty or sour odours, caking, chemical and nutritional changes, reduction in processing quality, and form of mycotoxins. These deteriorative changes affect the grade and price of grain and contribute to customer dissatisfaction when the grain is marketed. The respiration of grain and fungi results in a loss in dry matter as well as the production of heat and moisture which contribute to further spoilage. Net changes in nutritional value and the risk of mycotoxin contamination are difficult to predict because they depend on a complex interaction of factors such as temperature, moisture, storage time, fungal species composition, kind of grain, and previous storage history. Moisture is the most important variable determining the rate of deterioration caused by fungi, with temperature being the second major factor. Problems with deterioration of grain in export shipments are not a recent development. They are related primarily to grain moisture at loading, and also to the extent of previous mould invasion.     

Strategies to reduce mycotoxin levels in maize during storage: a review

Maize (Zea mays L.) is one of the main cereals as a source of food, forage and processed products for industry. World production is around 790 million tonnes of maize because as a staple food it provides more than one-third of the calories and proteins in some countries. Stored maize is a man-made ecosystem in which quality and nutritive changes occur because of interactions between physical, chemical and biological factors. Fungal spoilage and mycotoxin contamination are of major concern. Aspergillus and Fusarium species can infect maize pre-harvest, and mycotoxin contamination can increase if storage conditions are poorly managed. Prevention strategies to reduce the impact of mycotoxin in maize food and feed chains are based on using a hazard analysis critical control point systems (HACCP) approach. To reduce or prevent production of mycotoxins, drying should take place soon after harvest and as rapidly as feasible. The critical water content for safe storage corresponds to a water activity (a _w) of about 0.7. Problems in maintaining an adequately low a _w often occur in the tropics where high ambient humidity make the control of commodity moisture difficult. Damage grain is more prone to fungal invasion and, therefore, mycotoxin contamination. It is important to avoid damage before and during drying, and during storage. Drying maize on the cob before shelling is a very good practice. In storage, many insect species attack grain and the moisture that can accumulate from their activities provides ideal conditions for fungal activity. To avoid moisture and fungal contamination, it is essential that the numbers of insects in stored maize should be kept to a minimum. It is possible to control fungal growth in stored commodities by controlled atmospheres, preservatives or natural inhibitors. Studies using antioxidants, essential oils under different conditions of a _w, and temperature and controlled atmospheres have been evaluated as possible strategies for the reduction of fungal growth and mycotoxin (aflatoxins and fumonisins) in stored maize, but the cost of these treatments is likely to remain prohibitive for large-scale use.     

On ochratoxin A and fungal flora in Polish cereals from conventional and ecological farms - Part 1: occurrence of ochratoxin A and fungi in cereals in 1997.

Over 200 samples of Polish cereal grain from the 1997 harvest obtained from conventional and ecological farms were tested for the presence of ochratoxin A as well as for contamination by microscopic fungi. Ochratoxin A contamination of rye from ecological farms was over six times more frequent than that from conventional cultivation. The ochratoxin A content in wheat and barley samples from ecological farms was also higher. No wheat sample from conventional farms contained the mycotoxin. In the group of ecological farms, there were differences in the percentage of cereal samples containing ochratoxin A. The ochratoxin A levels ranged from 0.2 to 57 microg kg(-1). The mean concentration of ochratoxin A in investigated cereal grain was 5.7 microg kg(-1). From samples containing detectable amounts of ochratoxin A, fungi producing ochratoxin A under laboratory conditions were isolated. They were classified as belonging to the species Penicillium cyclopium, P. viridicatum, P. chrysogenum and also Aspergillus alliaceus, A. versicolor, A. glaucus and A. flavus. Penicillium strains - producers of ochratoxin A - were isolated from 93% of the samples; in 7% of samples, only Aspergillus strains producing this mycotoxin were noted. Rye samples mainly from one farm with an ecological type of cultivation and from one conventional farm were contaminated with both Aspergillus and Penicillium mycotoxigenic strains.     

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.     

Mycotoxins in cereal grain. Part VII. A simple method to assay mycotoxin potential of cereal grain and cereal products

A simple test to assay mycotoxin potential of cereal grain and products was elaborated. In cereal grain samples during 1977 and 1981 formation of ochratoxin A, citrinin and zearalenone was observed respectively in 40%, 7% and 31% of cereals samples. Aflatoxin and sterigmatocystin were not formed in any sample of cereal grain. Citrinin and penicillic acid were found as mycotoxins accompanying ochratoxin A.     

Natural Occurrence of Mycotoxins in Food and Feed: Pakistan Perspective

Fungi are commonly present in the environment and can grow under favorable conditions on an extensive variety of substrates. During harvesting, handling, storage, and distribution, agricultural commodities are subjected to infection by toxigenic molds, which may cause spoilage and produce toxic metabolites called mycotoxins. Fungal contamination of various food commodities with consequent exposure of the community to mycotoxins is a hazard that may exist depending on environmental factors, crop health, and soil conditions. Mycotoxins represent serious consequences due to substantial economic loss and risk to health. The environmental conditions of Pakistan with its mostly warm temperature are conducive to growth of toxigenic fungi resulting in mycotoxin production in different food items. Moreover, the poor conditions of storage and deficiency in regulatory measures in food quality control worsen the situation in the country. This review encompasses mycotoxin contamination of food and feed in Pakistan. High concentrations of mycotoxins are found in some commodities that are used on a daily basis in Pakistan, which may be a concern depending on dietary variety and health conditions of individuals in the population. Therefore, the mycotoxin contamination of foodstuff with exceeding levels represents a serious health hazard for the local population. There is a need to conduct more studies to analyze mycotoxin occurrence in all types of food commodities throughout the country. For consumer safety and the country's economy, the regulatory authorities should take into account this issue of contamination, and control strategies should be implemented and the quality control system of food improved.     

Mycotoxins in cereal grain Part VII. A simple method to assay mycotoxin potential of cereal grain and cereal products

A simple test to assay mycotoxin potential of cereal grain and products was elaborated. In cereal grain samples during 1977 and 1981 formation of ochratoxin A, citrinin and zearalenone was observed respectively in 40%, 7 % and 31 % of cereals samples. Aflatoxin and sterigmatocystin were not formed in any sample of cereal grain. Citrinin and penicillic acid were found as mycotoxins accompanying ochratoxin A.     

Ochratoxin A and citrinin loads in stored wheat grains: Impact of grain dust and possible prediction using ergosterol measurement

Crop storage should be carried out under hygienic conditions to ensure safe products, but sometimes grain dust which has settled from previous storage may be left over and incorporated to the following stored grains. This paper describes the results obtained using a lab model developed in order to assess the impact of grain dust incorporation for its direct contribution as a contaminant but also as an inoculum in stored wheat. Settled grain dust (4 samples) released from Belgian grain storages were collected and analysed by HPLC for ergosterol, ochratoxin A (OTA) and citrinin (CIT) content. For OTA and for ergosterol, there was a high degree of variability in concentrations found in the dust samples (from 17.3-318 ng g^-1 and from 39-823 µg g^-1, respectively) whilst for CIT, the range was less significant (from 137-344 ng g^-1). Incorporation of grain dust into wheat storage contributed to an increase in the concentrations of mycotoxins in the stored grain. Dust acts as a contaminant and as an inoculum. According to these two ways, patterns of mycotoxin generation vary with the nature of the mycotoxin, the mycotoxigenic potential of dust and the water activity of the wheat. OTA and CIT showed a very versatile image when considering the amounts of toxins produced under the selected experimental conditions. The development of a robust tool to forecast the mycotoxigenicity of dust was based on the determination of ergosterol content as a general marker of fungal biomass. Present results suggest that this predictive tool would only be valid for predicting the contamination level of CIT and OTA at reasonable moisture content (14-20%). The potential risk of having highly contaminated batches from stock to stock may thus occur and this paper discusses possible pathways leading to OTA and CIT contamination either under wet or dry storage conditions. We therefore, recommend taking precautionary measures not only by controlling and maintaining moisture at a reasonable level during storage of the raw materials but also by paying more attention to the cleaning of the stores before loading in the new harvests.     

干腌火腿中霉菌毒素研究的进展

从干腌火腿中分离到一些霉菌菌株在培养基中生长时可以产生黄曲霉毒素、环匹阿尼酸、青霉酸、梗曲霉素、展青霉素、灰黄霉素、霉酚酸、赭曲霉毒素、橘青霉素和疣孢苷啶等多种霉菌毒素。生成霉菌毒素的霉菌都属于青酶和曲霉。鲜绿青霉可以在干腌火腿中生成环匹阿尼酸,且这种霉菌毒素在干腌火腿中具有较高的稳定性,从而对干腌火腿的安全性构成很大的威胁。将无毒性的霉菌菌株进行培养并接种到干腌火腿上,可防止干腌火腿中形成霉菌毒素。     

Trichothecenes, ochratoxin A and zearalenone contamination and Fusarium infection in Finnish cereal samples in 1998

The occurrences and concentrations of trichothecenes, ochratoxin A and zearalenone in Finnish cereal samples are presented in this study. Furthermore, infections by moulds, especially Fusarium contamination of grains in the same samples, are reported. In total 68 cereal samples, including 43 rye, 4 wheat, 15 barley and 6 oats samples, were collected after a cool and very rainy growing season in 1998. A gas chromatograph combined with a mass spectrometric detector was used for determination of seven different trichothecenes. A high performance liquid chromatograph with a fluorescence detector was used for ochratoxin A and zearalenone determination. For the identification of moulds, the grain samples were incubated and the moulds were isolated and identified by microscopy. The analytical methods were validated for mycotoxin analysis and they were found to be adequately reliable and sensitive. Heavy rainfalls in the summer and autumn of 1998 caused abundant Fusarium mould infection in Finnish cereals, particularly in rye. Fusarium avenaceum was the most common Fusarium species found in cereals. However, the mycotoxin concentrations were very low and only deoxynivalenol, nivalenol and HT-2 toxin were detected. Deoxynivalenol was detected in 54 samples in the concentration range 5-111 µg/kg. Nivalenol and HT-2 toxin were detected in three and two samples, respectively, in the concentration range 10-20 µg/kg.     

Mould spoilage and mycotoxin formation in grains as controlled by physical means

Low oxygen concentrations (less than 1%) and/or increased concentrations of CO2 or N2 have been found to be highly effective in preventing the development of mould on grain and in inhibiting selected mycotoxins, e.g. aflatoxins, ochratoxin, patulin, penicillic acid and T-2. However, the levels of CO2 needed to inhibit mould growth are much higher than those required for the inhibition of mycotoxin production. The degree of inhibition achieved by elevated CO2 concentrations is dependent on other environmental factors, such as relative humidity (RH) and temperature. Nevertheless, the biosynthetic pathways for mycotoxin production are merely blocked, but not damaged by high CO2 levels. Irradiation has been shown to destroy the conidia of moulds but the information concerning the effect of irradiation on mycotoxin formation seems to be contradictory. Aflatoxin production was increased in irradiated wheat grain, but decreased in barley and maize when the grain was irradiated prior to inoculation. The number of spores in the inoculum, grain condition, relative humidity and other environmental factors could all affect the results obtained. However, ochratoxin formation by Aspergillus ochraceus was consistently enhanced by irradiation of spores or mycelium.     

Mycoflora and natural occurrence of aflatoxin, cyclopiazonic acid, fumonisin and ochratoxin A in dried figs

Mycoflora, the mycotoxigenic properties of moulds, and natural contamination with mycotoxins such as aflatoxins (AFs), cyclopiazonic acid (CPA), fumonisin B(1) (FB(1)) and ochratoxin A (OTA) were investigated in dried figs. Dry fig samples were collected from orchards during the drying stage in the Aegean Region of Turkey. Fungal isolates were identified using morphological, chemical as well as molecular methods. Mycotoxigenic characteristics of moulds were assessed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Mycotoxins except CPA (by TLC) were determined by HPLC. All the fig samples were contaminated with moulds and 94.7% contained one or more mycotoxigenic species. The most prevalent moulds present in dried figs belong to the Aspergillus section Nigri members, being 93.9% positive for the samples, followed by Fusarium spp., Aspergillus section Flavi and Penicillium spp. On the other hand, Fusarium spp. had the highest count and the number of fumonisin producing Fusarium was also high. A total of 48% of 115 dried fig samples contained OTA (range = 0.1-15.3 ng g(-1)), 74.7% of the samples had FB(1) (range = 0.05-3.65 mg kg(-1)), 10.0% of the samples had aflatoxin (range = 0.1-763.2 ng g(-1)) and 24.3% of the samples were tentatively identified as being contaminated with CPA (range = 25-187 ng g(-1)). Dried fig samples were contaminated with one (33.0%), two (47.0%), three (5.2%) and four mycotoxins (3.5%). A total of 11.3% of dried fig samples were not contaminated with any of the four mycotoxins. To the best of our knowledge, CPA and fumonisin have been found for the first time in dried figs.     

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.     

Mycotoxins in fruits,fruit juices,and dried fruits

This review gives an overview of the presence of mycotoxins in fruits. Although several mycotoxins occur in nature, very few (aflatoxins, ochratoxin A, patulin, Alternaria toxins) are regularly found in fruits. It has been shown that the presence of fungi on fruits is not necessarily associated with mycotoxin contamination. The formation of mycotoxins depends more on endogenous and environmental factors than fungal growth does. Mycotoxins may remain in fruits even when the fungal mycelium has been removed. Depending on the fruit and the mycotoxin, the diffusion of mycotoxins into the sound tissues of fruits may occur. The influence of the selection and storage of fruits and the influence of different processing steps involved in the production of fruit juices and dried fruits on possible mycotoxin contamination is described. It is shown that the careful selection, washing, and sorting of fruits is the most important factor in the reduction of mycotoxin contamination during the production of fruit juices. The processing of fruits does not result in the complete removal of mycotoxins.     

粮食真菌毒素污染的预防与脱毒

粮食真菌毒素的预防包括预防粮食作物田间生长及收获后储藏过程中毒素的生物合成及代谢。真菌毒素的脱毒主要指除去、破坏及减少毒素作用的收获后处理。田间及储藏中没能有效控制真菌毒素的合成必将导致对人类健康的危害及经济损失，而有效的监控将避免真菌毒素成为威胁人类健康的污染源。应用综合预防措施将是控制真菌毒素的有效策略。本文强调的收获前后措施将依特别年份的特定的气候条件而定。弄清适于真菌污染、生长和产毒环境因素是有效控制食物及饲料中真菌毒素的关键措施。有很多新的有效的收获前预防策略正在开发，如利用转基因技术创造粮食作物抗性新品种及利用非产毒真菌菌株生物防治等。收获后的防止真菌毒素产生主要依赖于收获前后的良好的管理措施。脱毒策略可分为物理、化学或微生物脱毒技术，这些脱毒技术主要通过破坏、修饰或吸附真菌毒素，从而达到减少或消除毒素作用。