Search

Mycotoxins produced by certain fungal species of the Fusarium genus are frequently found as contaminants in cereals and feedstuffs. Fumonisins (FBs), deoxynivalenol (DON) and zearalenone (ZEN) are of special concern relative to animal health and productivity. The aim of this work was to analyse the levels of Fusarium mycotoxin contamination in samples of total mixed rations (TMRs) for dairy cows. To accomplish this analysis, an HPLC-MS/MS multi-mycotoxin method was developed and validated. The relation between the formulation of TMR samples and the presence of mycotoxins was also studied. From February 2016 to January 2018, a total of 193 TMR samples for dairy cows collected from farms located in different areas of Spain were analysed for the presence of FBs, ZEN, DON and their metabolites. In total, 112 samples (58%) were contaminated with at least one mycotoxin, and 38 samples (20%) presented more than one mycotoxin. FBs were the mycotoxins most frequently found (34% positive samples). DON was detected in 17% of samples, and ZEN was detected in 16% of samples. Among the metabolites analysed, only deoxynivalenol-3-glucoside (DON-3-Glc) and 15-acetyldeoxynivalenol (15-ADON) were detected. The levels of all the Fusarium mycotoxins studied were always below the values recommended by the European Commission for feedstuffs. The wide variety of ingredients used in the formulation of the analysed samples made it difficult to reach definite conclusions, although it seemed that some cereal silages and concentrates such as cereals or compound feed used as ingredients of the TMR may be related to the presence of mycotoxins. This work was supported by the Spanish Ministry of Economy and Competitiveness (project AGL2014-55379-P).

Mycotoxins have several toxicological implications. In the present study, we evaluate the presence of aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin (FB1) in paddy rice, polished rice, and maize from the fields and markets in Nayarit State (Mexico). The results indicated the presence of AFB1 in 21.21% of paddy rice samples and 11.11% of market maize samples. OTA was present in only 3.03% (one sample) of paddy rice samples. FB1 was detected in 87.50% and 88.88% of maize samples from field and market, respectively. The estimated human exposure was calculated for FB1 using the probable daily intake (PDI), which suggested that FB1 could contribute to the development of diseases through the consumption of contaminated maize. Positive samples indicated that some rice and maize samples were not suitable for human consumption. Further efforts are needed to continue monitoring mycotoxins and update national legislation on mycotoxins accordingly This work was supported by the Consejo Nacional de Ciencia y Tecnolgía (CONACYT) under Grant code SALUD-2017-2-289726.

Cereal-based baby food production process is expected to have an impact on the initial level of Fusarium mycotoxins that can contaminate the raw materials. The aim of the present study was to investigate the changes of some of these toxins during roasting and the treatment with amylolytic enzymes, usually applied during the production process. Three different cereal flours contaminated with Fusarium graminearum were considered (barley, wheat and oat). The results did not show significant changes in the concentration of any of the studied mycotoxins (up to 5% change in deoxynivalenol concentration after the enzymes were added). The acetyl-deoxynivalenol also showed slight modifications as a result of the applied processes, however their statistical significance was not proved. Zearalenone and T-2 and HT-2 toxins remained almost unaltered throughout the study. The authors are grateful to the University of Lleida (grant JADE Plus 218/2016), and to the Spanish Ministry of Economy and Competitiveness (MINECO, Project AGL2017-87755-R) for funding this work.

A total of 64 samples of sorghum (37 Tunisian sorghum samples and 27 Egyptian sorghum samples) were collected during 2011 – 2012 from markets in Tunisia. Samples were analysed for contamination with aflatoxin B1, ochratoxin A and zearalenone by High-Performance Liquid Chromatography Coupled with Fluorescence Detection (HPLC-FLD). Aflatoxin B1 was found in 38 samples in the range 0.03 – 31.7 μg kg − 1 . Ochratoxin A was detected in 24 samples with concentrations ranging from 1.04 to 27.8 μg kg − 1 . Zearalenone was detected in 21 samples and the concentration varied between 3.7 and 64.5 μg kg − 1 . ANOVA analysis of the influence of the country of origin on the incidence and concentration of mycotoxins in the samples studied showed no significant difference ( P > 0.05) between the two batches of samples for each of the three mycotoxins studied. The studied mycotoxins contaminate sorghum and may also co-exist because of the diversity of the mycobiota in this cereal. The authors declare that there are no conflicts of interest and they are grateful to the Tunisian government for the financial support under Grant “ U12E503 ” .

Deoxynivalenol (DON) is one of the most frequently occurring mycotoxins in wheat crops worldwide and poses a risk to human and animal health due to its wide range of adverse effects. Deoxynivalenol-3-glucoside (DON-3-glucoside) is a DON plant conjugate that is widely found in cereal products. As DON accumulation in the field seems unavoidable, it is important to investigate all of the conditions that affect its stability during food processing. One of the most consumed cereal product around the world is bread, however the published information about DON stability in bread shows a large variability of results because a huge amount of factors affect DON and its modified forms. So, the aim of this research was to study the fate of DON and its modified forms through the breadmaking process with the addition of xylanase and α-amylase at different fermentation temperatures. Moreover, different α-amylase and xylanase concentrations were added to the dough to be fermented. To quantify DON and its derived forms in the samples, liquid chromatography with double mass spectrophotometer was used. DON was reduced during fermentation and baking; however, the reduction at each step was related to the fermentation temperature. The presence of α-amylase and xylanase caused increases in DON during fermentation and during early baking. DON-3-glucoside was slightly reduced after fermentation and was widely increased (>80%) after baking. Deepoxy-deoxynivalenol (DOM-1) increased during the breadmaking process. Breadmaking process can reduce DON concentration, however xylanase and α-amylase presence cause increases of DON. The authors are grateful to the Spanish government (project AGL2014-55379-P) for financial support. A. Vidal thanks the Spanish government (Ministry of Education) for the pre-doctoral grant.

Ochatoxin A (OTA) is one of the most important mycotoxins based on its toxicity. The oral route is the main gateway of entry of OTA into the human body, and specialized epithelial cells constitute the first barrier. The present study investigated the in vitro cytotoxic effect of OTA (5, 15 and 45 μM) and production of OTA metabolities in Caco-2 and HepG2 cells using a co-culture Transwell System to mimic the passage through the intestinal epithelium and hepatic metabolism. The results derived from MTS cell viability assays and transepithelial electrical resistance measurements showed that OTA was slightly cytotoxic at the lowest concentration at 3 h, but significant toxicity was observed at all concentrations at 24 h. OTA metabolites generated in this co-culture were ochratoxin B (OTB), OTA methyl ester, OTA ethyl ester and the OTA glutathione conjugate (OTA-GSH). OTA methyl ester was the major metabolite found in both Caco-2 and HepG2 cells after all treatments. Our results showed that OTA can cause cell damage through several mechanisms and that the OTA exposure time is more important that the dosage in in vitro studies. OTA methyl ester is proposed as an OTA exposure biomarker, although future studies should be conducted. The authors are grateful to the Spanish (Project AGL2011-24862) and Catalonian (XaRTA-Reference Network on Food Technology) Governments for their financial support. C.A. González-Arias thanks the Secretaria de Universitats i Recerca del Departament de Economia i Coneixement of the Generalitat de Catalunya for the pre-doctoral grant.

Polyamines are essential for all living organisms as they are involved in several vital cell functions. The biosynthetic pathway of polyamines and its regulation is well established and, in this sense, the ornithine descarboxylase (ODC) enzyme acts as one of the controlling factors of the entire pathway. In this work we assessed the inhibition of the ODC with D, l-α-difluoromethylornithine (DFMO) on Alternaria alternata and we observed that fungal growth and mycotoxin production were reduced. This inhibition was not completely restored by the addition of exogenous putrescine. Actually, increasing concentrations of putrescine on the growth media negatively affected mycotoxin production, which was corroborated by the downregulation of pksJ and altR, both genes involved in mycotoxin biosynthesis. We also studied the polyamine metabolism of A. alternata with the goal of finding new targets that compromise its growth and its mycotoxin production capacity. In this sense, we tested two different polyamine analogs, AMXT-2455 and AMXT-3016, and we observed that they partially controlled A. alternata's viability in vitro and in vivo using tomato plants. Finding strategies to design new fungicide substances is becoming a matter of interest as resistance problems are emerging. We thank Dr. Mark Burns, Aminex Therapeutics Inc. (USA), for kindly supply the polyamine analogs. We are also grateful to the Catalonian Government (XaRTA-Reference Network on Food Technology) for their financial support. N. Estiarte thanks the Secretaria de Universitats i Recerca del Departament d’Economia i Coneixement of the Generalitat de Catalunya for the predoctoral grant (FI-DGR 2013).

The objective of the present study was to assess the differences in modeled growth/AFB1 production probability and kinetic growth parameters for Aspergillus flavus inoculated as single spores or in a concentrated inoculation point (~500 spores). The experiment was carried out at 25°C and at two water activities (0.85 and 0.87) on pistachio extract agar (3%). Binary data obtained from growth and AFB1 studies were modeled using linear logistic regression analysis. The radial growth curve for each colony was fitted to a linear model for the estimation of the lag phase for growth and the mycelial growth rate. In general, radial growth rate and lag phase for growth were not normally distributed and both of them were affected by the inoculation type, with the lag phase for growth being more affected. Changing from the multiple spore to the single spore inoculation led to a delay of approximately 3-5days on the lag phase and higher growth rates for the multiple spore experiment were found. The same trend was observed on the probability models, with lower predicted probabilities when colonies came up from single spores, for both growth and AFB1 production probabilities. Comparing both types of models, it was concluded that a clear overestimation of the lag phase for growth occurred using the linear model, but only in the multiple spore experiment. Multiple spore inoculum gave very similar estimated time to reach some set probabilities (t10, t50 and t100) for growth or AFB1 production due to the abruptness of the logistic curve developed. The observed differences suggest that inoculum concentration greatly affects the outcome of the predictive models, the estimated times to growth/AFB1 production being much earlier for the concentrated inoculum than for a single spore colony (up to 9days). Thus the number of spores used to generate data in predictive mycology experiments should be carefully controlled in order to predict as accurately as possible the fungal behavior in a foodstuff.

The predominant fungal species present in 60 samples of barley collected in Spain were Alternaria alternata , Penicillium spp. and Aspergillus flavus . Of the 176 Alternaria isolates examined, 88.6% produced tenuazonic acid, 15.3% produced alternariol, and 9% produced alternariol monomethyl ether. Only 6% of the 190 isolates of A. flavus produced aflatoxin.

Malting was simulated using two different batches of barley as raw material: a naturally contaminated batch and laboratory inoculated (with a deoxynivalenol (DON) and zearalenone (ZEA) producing Fusarium graminearum strain) one. Up to three contamination levels were prepared, every process being carried out in triplicate. A significant washout effect on DON was observed by the end of the first water phase (between 22.4 and 34% reduction) with an even more pronounced reduction (up to 75% decrease) by the end of the steeping process. ZEA content remained almost unchanged (no significant difference between the initial and the final concentration). Germination was characterized by an increase in all the three toxins (ZEA, DON and DON-3-G) concentrations, however showing a decreasing trend in the last 24 h of the stage, compared to the first day of germination. Kilning lead to a significant reduction of DON in the naturally contaminated batch (46.6 and 78.8%), nevertheless an increase in all other toxins and contamination levels was observed. The authors are grateful to the University of Lleida (grant JADE Plus 218/2016), and to the Spanish Ministry of Economy and Competitiveness (MINECO, Project AGL2014-55379-P) for funding this work. The authors are also grateful to the malting plant Malteria la Moravia SL for the supplied material and installations.

Silages constitute a major component of the feed ration for dairy cows, being a potential source of mycotoxins due to the possible contamination by filamentous fungi capable of producing these toxic compounds. In this study, samples of different kinds of silages collected from farms located in four regions of Spain, were analysed to evaluate the occurrence of aflatoxins (AFs) and Fusarium mycotoxins. Lactic acid bacteria and fungal populations as well as pH and water activity were also studied. Penicillium, Geotrichum and Monascus were the main fungi identified in all the silages examined. The incidence of AFs was low (10 % of positive samples). Fusarium mycotoxins were detected in 40 % of the samples and fumonisins (FBs) were the most commonly detected. Maize silage was the most heavily contaminated type of silage. Levels of mycotoxins detected in positive samples did not exceed the EU guidance values. The lack of relationship between Fusarium counts and its mycotoxin concentrations suggested that mycotoxin production possibly occurred pre-ensiling or immediately post-ensiling. Outcomes showed that mould growth and mycotoxin contamination in silages should be regularly monitored in order to minimize the exposure of dairy cows to contaminated feed. This work was supported by the Spanish Ministry of Economy and Competitiveness (project AGL2014-55379-P). María Rodríguez-Blanco thanks the University of Lleida for a pre-doctoral grant. The authors would like to thank Montserrat Prim for her technical assistance.

The stability of deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON-3-glucoside) during the breadmaking process was studied. Some enzymes used in the bakery industry were examined to evaluate their effects on DON and DON-3-glucoside. The level of DON in breads without added enzymes was reduced (17-21%). Similarly, the addition of cellulase, protease, lipase and glucose-oxidase did not modify this decreasing trend. The effect of xylanase and alpha-amylase on DON content depended on the fermentation temperature. These enzymes reduced the DON content by 10-14% at 45 degrees C. In contrast, at 30 degrees C, these enzymes increased the DON content by 13-23%. DON-3-glucoside levels decreased at the end of fermentation, with a final reduction of 19-48% when no enzymes were used. However, the presence of xylanase, alpha-amylase, cellulase and lipase resulted in bread with greater quantities of DON-3-glucoside when fermentation occurred at 30 degrees C. The results showed that wheat bran and flour may contain hidden DON that may be enzymatically released during the breadmaking process when the fermentation temperature is close to 30 degrees C. The authors are grateful to the Spanish government (project AGL2014-55379-P) for financial support. A. Vidal thanks the Spanish government (Ministry of Education) for the pre-doctoral grant.

Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37 ◦C), water activity (aw, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 aw at 37 ◦C for two of the isolates. The minimum aw needed for mycelial growth was 0.91 at 25 and 37 ◦C. At 15 ◦C, only isolate 8 grew at 0.99 aw. Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 aw). Aflatoxin production was not observed at 15 ◦C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains. We thank M. Prim for her encouragement and technicalassistance. The authors are grateful to the European Union(MYCORED KBBE-2007-2-5-05 project) and Tunisian Govern-ment for financial support.

Deoxynivalenol (DON) is a mycotoxin produced mainly by Fusarium species and occurs predominantly in cereal grains such as wheat. Due to its toxic effects, in the European Union DON content in unprocessed cereals and processed cereal-based products for human consumption has been regulated, and recommended maximum limits have been established for animal feed. In this study, a method for degrading DON on wheat kernels, by exposition to ammonia (NH3) vapours, was optimized. Results have shown that with a simple treatment with ammonia vapours at 90ºC (for two hours), degradations higher than 75% were achieved in kernels affected by a moderated contamination up to 2000 µg/kg DON. The study of the reaction between DON and NH3 allowed us to tentatively establish the structure of possible degradation products. In addition, in silico evaluation indicated, in general, lower toxicity and biological effects for the degradation products than for DON. B.B.-V. and J. J R.-B. appreciate the FD pre-doctoral fellowship (PRE2018-085278) of the Spanish Ministry of Science, Innovation and Universities, and the postdoctoral contract from Agrotecnio center, respectively. This work was funded through the project AGL2017-87755-R awarded by the Spanish Ministry of Science, Innovation and Universities.

Hyperspectral imaging (HSI) is an emergent, rapid, cost-effective and non-destructive technique in which the spectral data are obtained for each pixel location in a sample's image. The application of this technique to assess mycotoxins and mycotoxigenic fungi in cereals is considered promising to replace time-consuming wet-chem- istry methods and for its potential grain sorting ability, in order to reduce food and feed contamination and the associated toxic effects in human and animals. Fusarium is a plant pathogen and deoxynivalenol (DON)-producer which presents high incidence in cereals such as wheat, maize and barley. The following review encompasses detailed information about the HSI principle and an updated outlook of its applications in the detection and quantification of Fusarium and DON in cereals. Moreover, HSI prediction algorithms for DON quantification are novel approaches which present high complexity owed to the asymptomatic nature of the grain despite of high mycotoxin concentrations. The spatial faculty of this system may be able to overcome the contamination het- erogeneity of the grain for its elimination, enhancing risk management and rising the economic performance. Additionally, HSI is also proposed as a powerful grain sorting instrument due to high accuracies obtained in classification of single kernels according to Fusarium and DON infection. Therefore, an overview of the HSI applications for on-line and massive cereal sorting in grain industry is also presented. The authors are grateful to the University of Lleida (predoctoral grant), and to the Spanish Ministry of Science, Innovation and Universities (Project AGL2017-87755-R) for funding this work.

Mycotoxins are secondary metabolites produced by fungi that may contaminate different foods intended for human consumption, resulting in a widespread exposure worldwide. The novel exposome paradigm has the ambition to decipher the different environmental insults threating human health throughout the entire lifespan. Given the large potential impact of mycotoxins in terms of human exposure and related health effects, the ambition of this review is to present this group of chemical compounds and the high interest to be included in exposome projects. Furthermore, we also attempt to approach the novel exposome paradigm to more traditional disciplines such as mycotoxin exposure assessment and mycotoxicology, introducing the new methodological challenges and translational needs. Hence, we provide an overview of major biomarkers currently developed, biological matrices where these may be found, an overview of internal exposure levels and potential co-occurrence with environmental chemicals and finally an overview of major health effects with the illustrative example of the potent xenoestrogen zearalenol. Conversely, these new approaches may be an excellent opportunity to fill many research gaps on mycotoxins research as the identification of associations with human health, elucidation of join effect with other environmental exposures or the decipher of underlying molecular mechanisms by using advanced OMICs technologies. The authors are grateful to the Spanish Ministry of Science, Innovation and Universities (Project AGL2017- 87755-R) for funding this work.

Review Contamination of animal feed with mycotoxins still occurs very often, despite great efforts in preventing it. Animal feeds are contaminated, at low levels, with several mycotoxins, particularly with those produced by Aspergillus and Fusarium genera (Aflatoxin B1, Ochratoxin A, Zearalenone, Deoxynivalenol and Fumonisina B1). In animal feed, to date, only Aflatoxin B1 is limited through EU regulation. Consequently, mycotoxins cause serious disorders and diseases in farm animals. In 2009, the European Union (386/2009/EC) approved the use of mycotoxin-detoxifying agents, as feed additives, to prevent mycotoxicoses in farm animals. The present review gives an overview of the problem of multi-mycotoxin contamination of feed, and aims to classify mycotoxin adsorbing agents (minerals, organic, and synthetic) for feed decontamination, focusing on adsorbents with the ability to bind to multiple mycotoxins, which should have a more effective application in farms but they are still little studied in scientific literature. The authors are grateful to the Spanish government (projects RTC-20153508-2 and AGL2014-52648-REDT) and to XaRTA (The Catalonian Reference Network in Food Technology) for funding. They also would like to thank Arthur Kendall for editing the English usage in the manuscript. P. Vila-Donat thanks MINECO and the University of Lleida for her postdoctoral contract.

A multianalyte method, using a MS/MS detector, was applied for a simultaneous determination of 23 mycotoxins in 64 beer products purchased from the supermarket in Lleida, Spain. The samples varied by their origin, brewing technology, alcohol content, etc. The results showed that 20.3% of the tested samples were mycotoxin contaminated overpassing the limit of detection (LOD). None of the alcohol-free samples (17%) were contaminated with mycotoxins. The most frequently occurring toxin was zearalenone (ZEN), being quantified in 65% of the positive samples, with levels ranging from 8.24 to 62.96 μg/L. Regarding the co-occurrence of mycotoxins, three samples were found to contain two or more mycotoxins simultaneously. A deterministic approach was used to evaluate the contribution of beer consumption to daily intake and the proportion of the established tolerable daily intake (TDI) for ZEN and deoxynivalenol (DON) and its metabolite deoxynivalenol-3-glucoside (DON-3-G). The authors are grateful to the University of Lleida (grant JADE Plus 218/2016), and to the Spanish Ministry of Economy and Competitiveness (MINECO, Project AGL2014-55379-P) for funding this work.

The objective of this study was to characterize the mycoflora of sorghum grains commercialized in the Tunisian retail market and to identify aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) producing species. Sixty four samples of sorghum (37 samples of Tunisian sorghum and 27 samples of Egyptian sorghum) were analyzed. Dilution plating (CFU, colony forming units) was used for fungal enumeration. The isolation of mycobiota was carried out by plating of grains on PDA and malachite green medium. Aspergillus section Flavi and section Nigri and Fusarium isolates were sub-cultured in CYA to test their ability to produce AFs, OTA and ZEA, respectively. The selected Aspergillus section Flavi and section Nigri, Penicillium and Fusarium isolates were subjected to specific PCR assays using published species-specific primers. The results revealed the dominance of Fusarium (95.3%), followed by Aspergillus (87.2%) and Alternaria (81.2%) species. The fungal counts ranged from 100 to 1.3·104 CFU/g for Tunisian sorghum and from 100 to 5.7·103 CFU/g for Egyptian sorghum. Among Aspergillus section Flavi isolates identified by molecular biology, Aspergillus flavus was the most dominant (90.1%) while Aspergillus parasiticus represent 9.9% only. About Aspergillus section Nigri, results showed the dominance of Aspergillus niger aggregate species, including Aspergillus niger, Aspergillus tubingensis and other species. Among Fusarium species, Fusarium incarnatum was the most dominant in both Tunisian and Egyptian sorghum. Penicillium citrinum was the dominant Penicillium species in the studied samples. More than 890 isolates belonging to the genus Aspergillus and Fusarium were tested in order to test their capacity to produce AFs, OTA and ZEA. The percentage of mycotoxin producing isolates in Aspergillus section Flavi, A. section Nigri, and Fusarium was 30.0%, 4.6% and 11.1%, respectively. We thank Montse Prim for her encouragement and technical assistance. The authors are grateful to the European Union (MYCORED KBBE-2007-2-5-05 project) and Tunisian government for the financial support.

The effects of two exposure times per day (6 and 16 h) of UV-A or UV-B radiation, combined with dark and dark plus light incubation periods during 7e21 d on fungal growth and mycotoxins production of Aspergillus species were studied. Aspergillus carbonarius and Aspergillus parasiticus were inoculated on grape and pistachio media under diurnal and nocturnal temperatures choosing light photoperiod according to harvest conditions of these crops in Spain. Ultraviolet irradiation had a significant effect on A. carbonarius and A. parasiticus colony size (diameter, biomass dry weight, and colony density) and mycotoxin accumulation, although intraspecies differences were observed. Inhibition of A. carbonarius fungal growth decreased when exposure time was reduced from 16 h to 6 h, but this was not always true for ochratoxin A (OTA) production. OTA reduction was higher under UV-A than UV-B radiation and the reduction increased along time conversely to the aflatoxins (AFs). Aflatoxin B1 (AFB1) was the main toxin produced by A. parasiticus except in the UV-B light irradiated colonies which showed a higher percentage of AFG than AFB. Morphological changes were observed in colonies grown under UV-B light. The authors are grateful to the Spanish Government (AGL2010-22182-C04-01) and Comissionat per a Universitats i Recerca del Departament d’Innovació, Universitats i Empresa de la Generalitat de Catalunya (AGAUR). The authors are also grateful to Fernando Rubio and Hector Castan by the laboratory support.

Aflatoxins are the most potent genotoxic and carcinogenic mycotoxins. To date, research has only focused on the presence of free aflatoxins in agricultural commodities. Therefore, the main objective of this study was to investigate the occurrence of possible modified aflatoxins in maize. Different hydrolysis methods were applied to convert modified mycotoxins into their free aflatoxins. Eighteen aflatoxin-contaminated maize samples were incubated with potassium hydroxide, trifluoromethanesulfonic acid and several enzymes to induce hydrolysis. Potassium hydroxide caused a total reduction of aflatoxins, while trifluoromethanesulfonic acid did not lead to an increase in free aflatoxins, neither did treatment with a protease. However, α-amylase and cellulase incubation caused significant increases in the total free aflatoxin content, 15 ± 8% and 13 ± 5%, respectively. These results show that a small proportion of aflatoxins could be associated to matrix substances in plants. Consequently, hydrolysis could occur during food processing and during mammalian digestion, leading to an underestimation of the total aflatoxin content. The authors acknowledge the Spanish government (project AGL2014-55379-P) for the financial support. A. Vidal acknowledges the Spanish government (Ministry of Education) for the pre-doctoral grant. The authors are grateful to Mrs. Cynthia Chilaka for the kind provision of the maize samples.

The aim of this study was to evaluate the effect of 15 commercial yeasts in the mitigation of the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) during the brewing process. Saccharomyces strains (10 strains of S. cerevisiae and 5 of S. pastorianus) were used to ferment DON and ZEN contaminated wort. Wort samples were taken every 24 h during fermentation, while mycotoxin analysis in yeast was performed at the end of fermentation (96 h); additionally, pH and ethanol content were measured daily. For mycotoxin analysis, after immunoaffinity purification of sample extracts, analysis was performed using an Ultra-High-Pressure Liquid Chromatograph coupled with a diode array or fluorescence detector (UHPLC-DAD/FLD). Mycotoxin presence had no significant effect on the ethanol production during brewing. At the end of fermentation, 10-17% of DON and 30-70% of ZEN had been removed, 6% of the initial concentration of DON and 31% of the ZEN being adsorbed by the yeast. Beermakers must pay careful attention to the raw material since a high percentage of DON could be present at the end of the beer fermentation process. Future studies should focus on the quantification of "masked" mycotoxins that are relevant to food security. The authors are grateful to the University of Lleida (JADE Plus 218/2016 grant), to the Spanish Ministry of Economy and Competitiveness (MINECO, AGL2014-55379-P project) for funding this work. Wall-Martínez HA is grateful to the Consejo Nacional de Ciencia y Tecnología(CONACyT) of Mexico for the scholarship.

A survey of fungal contamination and presence of aflatoxins (AFs) and fumonisins (FBs) in 30 feed samples collected from 10 tilapia farms during three seasons in Nayarit State, located in north-western Mexico, was carried out using ELISA as screening and High-Performance Liquid Chromatography (HPLC) as confirmatory method. Mycobiota included Aspergillus flavus and Fusarium spp. AFs were detected in 63.3% of samples using ELISA, but confirmation by HPLC revealed that all samples were under the detection limit. Regarding to FBs, positive samples were detected using both methods, with 19 positive samples (60% of total) by ELISA and 14 positive samples (46.6% of total) by HPLC and levels ranging from 0.148 to 2.587 mg/kg. Correlation was observed between both methods (r = 0.516, p = 0.004) for FBs results. No sample exceeded the European maximum levels for any of the mycotoxins. Water activity of samples ranged from 0.345 to 0.655, suggesting that mycotoxin occurrence is probably related to raw material conta...

Eighty one (81) samples of durum wheat collected from seven areas from Morocco (Khmiset, Beni Mellal, Settat, Fes–Meknes, Skhirat–Temara, Tansift-El Haouz and Rabat–Sale) were surveyed for the presence of the mycotoxin deoxynivalenol (DON). Samples were extracted with water, the extracts were purified by immunoaffinity chromatography (IAC) columns and DON was then identified and quantified with liquid chromatography (LC) coupled to diode array detection (DAD). Analytical results showed that 9 out of 81 total samples (11.1%) were contaminated with DON. DON levels in positive samples ranged between 65 and 1310 μg/kg. The maximum contamination level of DON (1310 μg/kg) was found in a durum wheat sample from Rabat–Sale area, while the minimum value (65 μg/kg) was registered in a sample from Beni Mellal area. The mean DON level in positive samples was 502.1 ± 40.4 μg/kg. Only one previous work exists on DON contamination in Moroccan wheat samples.

The present study aimed to assess the effect of some pre-harvest fungicides commonly used in Capsicum fruit production on growth and ochratoxin A production of three Aspergillus species found in Capsicum powder. Aspergillus tubingensis, Aspergillus ochraceus and Aspergillus westerdijkiae, previously isolated from paprika and chilli, were inoculated on yeast extract sucrose agar and paprika extract agar supplemented with different fungicides at their recommended dosage rates, and incubated at 20 and 30 °C during 7 days. Radial growth was measured after 3, 5 and 7 days and ochratoxin A production was determined by high-performance liquid chromatography with fluorescence detection on day 7. Dodine 40% and mancozeb 80% were the most effective fungicides in inhibiting growth and ochratoxin A production, regardless of the fungal strain tested or temperature conditions. Whereas the application of fungicides could be very attractive in reducing the mycotoxigenic fungal load, it can also stimulate ochratoxin A production in some cases.

Predictive mycology has dealt mainly with germination, growth and inactivation of fungi while the issue of mycotoxin production remains relatively unexplored. Very few studies provide biomass dry weight/colony size data along with mycotoxin data for the same sample times, thus the ratio mycotoxin accumulation per fungal biomass dry weight/colony size has rarely been reported. For this reason, the objective of the present study was to model the kinetics of mycotoxin production under the assumption of existing both no-growth-associated and growth-associated production. Aspergillus flavus was chosen as a model mycotoxigenic microorganism, and it was grown in maize agar medium and maize grain at 0.90 and 0.99 aw at 25 °C. A significant positive correlation (p < 0.05) was observed among the biomass responses (colony radius and biomass dry weight) in agar medium and colony radius in maize at both aw levels assayed. The Luedeking–Piret model was used to model AFB1 production and reasonable percentages of variability were explained. Moreover, AFB1 production was in general slightly better predicted through colony area. As conclusion, aflatoxin production may follow a mixed-growth associated trend, confirming that toxin formation does not present a clear delay in relation to growth under certain conditions.

BACKGROUND: Maize is a very important cereal for human and animal diet, but it can be contaminated by moulds and their mycotoxins. On the other hand, natural plant products with antimicrobial properties could possibly used to control mycotoxigenic fungi in foods and feeds. In this study, Equisetum arvense extract was tested for the efficacy on Aspergillus section Flavi and Fusarium section Liseola growth. Natural contaminated maize was used in this study and extract was added under different water activities (aw) – 0.90 and 0.95 – for Aspergillus section Flavi and Fusarium section Liseola, respectively. Moulds were inoculated in maize and incubated during 30 days. RESULTS: We confirm that E. arvense extract may be effective for the inhibition of Aspergillus section Flavi in maize with high levels of this mould. Moreover, this extract showed a good inhibition of growth on Fusarium section Liseola levels. Aflatoxin and fumonisin production was not affected by the extract. CONCLUSIONS: E. arvense extract could be an alternative to synthetic fungicides to control maize mycobiota level in moist grain. c � 2012 Society of Chemical Industry

Available information on the prediction of postharvest production of mycotoxins in recent years is reviewed. Predictive mycology has been focused mainly on fungal growth whereas studies on prediction of mycotoxins in foods are scarce. Modeling mycotoxin production is challenging due to the high variability in mycotoxigenic potential among species and isolates. Besides mycotoxin biosynthesis pathways and factors influencing them are still poorly understood. Baranyi and Luedeking-Piret models have been recently used as primary models for mycotoxin prediction, while for secondary modeling, polynomial approaches have been used. Furthermore, probability models can be a different alternative. In any case, media for data generation, intraspecies variability, and microbial interactions should not be disregarded before model application in food safety management systems. The authors are grateful to the Agència de Gestió d’Ajuts Universitaris i de Recerca de la Generalitat de Catalunya (AGAUR, grant 2014FI_B 00045), and to the Spanish Ministry of Economy and Competitiveness (MINECO, Project AGL2014-55379-P) for funding this work.

Novel strategies that address vitamin A deficiency have been developed, such as high-carotenoid maize, a biofortified transgenic maize line rich in carotenoids generated by genetic transformation. The South African white maize inbred (M37W), which is devoid of carotenoids, was engineered to accumulate high levels of β-carotene (provitamin A), lutein, and zeaxanthin. Maize seeds contaminated with fumonisins and other mycotoxins pose a serious threat to both humans and livestock. During three consecutive harvests, the fungal incidence and the fumonisin and aflatoxin content of maize seeds grown in an experimental field in Catalonia (Northeastern Spain) were evaluated. Fungal infection was similar in high-carotenoid maize and its isogenic line, with Fusarium verticillioides being the most prevalent fungus in all the harvests. Neither Aspergillus spp. nor aflatoxin contamination was found. Fumonisin levels were lower in high carotenoid than in its isogenic line, but this reduction was statistically significant in only 2 of the 3 years of study. Our results suggest that high carotenoid content reduces fumonisin levels in maize grains. The study was supported by La Caixa (Recercaixa project PC084082 VitaMaize: high-quality and safe food through antioxidant fortified maize), the Agrotecnio Center, the Catalonian Government (2014 SGR 1296 Agricultural Biotechnology Research Group and XaRTA Reference Network on Food Technology) and the Spanish Ministry of Economy and Competitiveness (BIO2014-54441-P). J. Díaz-Gómez thanks the University of Lleida for a pre-doctoral grant. The authors would like to thank the Agricultural Biotechnology group of the University of Lleida, Antonio Michelena, and Jaume Lloveras for their work in the experimental field.

Ethylene is a naturally produced plant regulator involved in several plant functions, such as regulation of fruit ripening. Inhibition of ethylene perception by using 1-methylcyclopropene (1-MCP) slows down the ripening of the fruit maintaining its quality and freshness. The use of 1-MCP is a commercial strategy commonly used in the food industry to extend the postharvest life of several fruits, including tomatoes. To assess how 1-MCP affected infection by Alternaria alternata on tomatoes, three different cultivars were artificially inoculated with 5 μL of an A. alternata conidial suspension (105 conidia/mL). Tomatoeswere treatedwith 0.6 μL/L of 1-MCP for 24 h. Spiked but untreated tomatoes were considered controls. Then, fruit were stored 6 days at 10 °C and one more week at 20 °C to simulate shelf-life. Fungal growth development and mycotoxin production (alternariol, AOH and alternariol monomethyl ether, AME) were assessed both on the first and on the second week. After the first 6 days at 10 °C, in just one variety the black mold disease was higher in the 1-MCP treated samples. However, after two weeks of storage, in all cases, tomatoes treated with 1-MCP showed more significant fungal growth disease. Regarding mycotoxin production, no large differences were observed among different treatments, which was corroborated with gene expression analysis of pksJ, a gene related to AOH and AME biosynthesis. We thank the Postharvest Programme of IRTA, especially to J. Usall, E. Costa and R. Altisent, for kindly offering us their postharvest chambers and laboratories and for helping us with the fruit quality measurements. We are also grateful to the Catalonian Government (FI-DGR 2013) (XaRTA-Reference Network on Food Technology) for their financial support. N. Estiarte thanks the Secretaria de Universitats i Recerca del Departament d'Economia i Coneixement of the Generalitat de Catalunya for the pre-doctoral grant.

The mycotoxins aflatoxins (AFs), deoxynivalenol (DON), fumonisins (FBs) and zearalenone (ZEA) are produced by fungal species can contaminate cereal-based food and therefore can be found alone or simultaneously in cereal-based commodities. The objectives of this study were, to study the occurrence of AFs, DON, FBs and ZEA in ethnic and gluten-free food samples from Lleida (Spain), and also to assess the exposure of immigrant and celiac population to the selected mycotoxins. Ethnic food and gluten-free samples were purchased in specialized grocery stores from Lleida (Spain). Mycotoxins were extracted and purified from the samples using immunoaffinity chromatography (IAC) columns. Determination and quantification of AFs, ZEA and FBs was performed with high performance liquid chromatography (HPLC) with fluorescence detector, and coupled to absorbance detector in case of DON. The results showed that FBs were widely found in corn-based foods purchased from special shops specialized on imported foods. DON was mainly found in cuscus and several corn-based ethnic foods. Several gluten-free food samples were contaminated by FBs (6 out of 18) and DON (2 out 18), while AFs and ZEA only were found in 6 ethnic foods samples. Considering these results, these special commodities should not be a health concern for these collectives; however, special attention should be focused on bulk imported foods distributed in special retail shops.

BACKGROUND:Fusariumgraminearum andF.verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans. RESULTS: F.graminearum grew better when was incubated at 15, 20 and 15 – 20 ◦ C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µ gg −1 , respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 ◦ Cf or zearalenone and 20 ◦ C for deoxynivalenol. F.verticillioides grew better at 25 ◦ C in culture medium and at 15/20 ◦ C and 15/25 ◦ C on soybean seeds. Fumonisin B1 was produced only in culture medium, and the maximum level (7.38 µ gg −1 ) was found at 15 ◦ C after 7 days of incubation. CONCLUSION: When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions. c � 2012 Society of Chemical Industry

HT-2 toxin (HT2) and T-2 toxin (T2) are mycotoxins produced in several cereals by species of Fusarium. The aim of the present study was to assess the exposure of Catalonian population to the combined mycotoxins T2 and HT2. Three different approaches were considered to handle the left censored data: (1) a substitution method, (2) a parametric method using the maximum likelihood estimation (MLE) and (3) a non-parametric method using the Kaplan-Meier (KM) estimator. Raw contamination data was provided by a large survey conducted in this region, in addition to the raw consumption data from a nutritional study specifically designed to assess the dietary intake of the main foodstuffs related to T2 and HT2 contamination for all population age groups. Contamination and consumption data were combined by simulation using a stochastic method. Accuracy and reliability of the statistic estimates were assessed building the related confidence intervals using a pseudo-parametric bootstrap method. Considering the results from the present study, the most exposed group population to the toxin T2 and HT2 should be expected to be the children.

Aflatoxins (AFs) are the main contaminants in pistachio nuts. AFs production in pistachio has been attributed to Aspergillus flavus. The aim of this study was to apply existing models to predict growth and AFs production by an A. flavus isolated from pistachios as a function of moisture content and storage temperature of pistachios in order to test their usefulness and complementarities. A full factorial design was used: the moisture content levels assayed were 10, 15, 20, 25 and 30% and incubation temperatures were 10, 15, 20, 25, 30, 37 and 42 °C. Both kinetic and probability models were built to predict growth of the strain under the assayed conditions. Among the assayed models, cardinal ones gave a good quality fit for radial growth rate data. Moreover, the progressive approach, which was developed based on a reduced number of experimental points led to an improved prediction in the validation step. This is quite significant as may allow for improved experimental designs, less costly than full factorial ones. Probability model proved to be concordant in 91% of the calibration set observations. Even though the validation set included conditions around the growth/no-growth interface, there was a 100% agreement in the predictions from the data set (n = 16, cut off = 0.5) after 60 days. Similarly, the probability for AF presence was rightly predicted in 89% of the cases. According to our results EC maximum aflatoxin levels would be surpassed in a period as short as 1 month if pistachio nuts reach 20 °C, unless %mc is ≤10%. The authors are grateful to Spanish government (project AGL2010-22182-C04-04) and EC, KBBE – Food, Agriculture and Fisheries and Biotechnology (project 222738- Selection and improving of fit-for-purpose sampling procedures for specific foods and risks), for their financial support.

Penicillium expansum is the main species responsible for patulin production in apples and pears. Generally, fruit is stored at suboptimal conditions for mould growth and this situation could influence on the intra-species variability in both capability for growth and mycotoxin production. The aim of this research was to assess the impact of suboptimal environmental conditions on the intra-specific variability of P. expansum growth and patulin production using seventy nine isolates of this mould. Petri dishes with Apple Concentrate Agar Medium (ACAM) were inoculated centrally and incubated at two temperatures, one near optimal (20 °C) and the other representative of suboptimal cold storage (1 °C). For each condition, 10 Petri dishes were inoculated, and colony growth and patulin production was measured over time. The Kruskal–Wallis test revealed significant differences among growth rate (μ) and lag phase (λ) within the seventy nine assayed isolates. Coefficients of variation revealed a wider dispersion of μ (mm/day) and λ (days) at 1 °C compared with 20 °C. There were significant differences (p < 0.05) among patulin levels (ng/mm2) for the different conditions, values being lower at the lower temperature. Coefficients of variation revealed a wider dispersion of mycotoxin production at 1 °C. In order to address the strain variability in growth initiation and prove the well-established notion of reducing patulin production in foods by preventing fungal growth, a greater number of strains should be included. The authors are grateful to the European (MYCOREDKBBE-2007-2-5-05 project), Spanish (AGL2010-22182-C04-04 project), Comissionat per a Universitats i Recerca d'Innovació, Universitats I Empresa de la Generalitat de Catalunya (AGAUR) and European Social Fund for the financial support.

This paper presents a journey through the known and the presumed history of two important mycotoxicoses which played an important role in the history of mankind before aflatoxins were discovered: (1) ergotism, also known as St. Anthony's fire or Holy Fire, linked to the consumption of cereals (especially rye) contaminated by the hallucinogenic and vasoconstrictor alkaloids produced by the Claviceps purpurea ergot, whose presence in Europe during the Middle Ages was considered epidemic, and (2) the so-called alimentary toxic aleukia (also known by its English acronym ATA), caused by the trichothecenes produced by Fusarium sporotrichioides and Fusarium poae, which devastated a large part of Russia shortly before the characterisation of the toxins of Aspergillus flavus.

The aim of this study was to assess the impact of suboptimal environmental conditions on the intraspecific variability of A. carbonarius growth and OTA production using thirty isolates of A. carbonarius. Three a(w)/temperature conditions were tested, one optimal (0.98a(w)/25°C) and two suboptimal: 0.90a(w)/25°C and 0.98a(w)/37°C as suboptimal water activity and temperature, respectively, which might take place through over ripening and dehydration of grapes. For each condition, 12 Petri dishes were inoculated, and colony growth and OTA production were measured over time. ANOVA revealed significant differences among μ and λ within the 30 assayed isolates. Coefficients of variation (CV%) revealed a wider dispersion of growth rates at 0.90a(w)/25°C compared to 0.98a(w)/25°C, and a more than 4-fold higher CV at 0.98a(w)/37°C compared to 0.98a(w)/25°C. However, dispersion of lag phases was similar at 0.98a(w)/25°C and 0.90a(w)/25°C and wider at 0.98a(w)/37°C. There were significant differences (p0.05) among OTA levels (ng/mm(2)) for the different conditions, values being lower under marginal conditions, and particularly at 0.98a(w)/37°C. Coefficients of variation (CV%) revealed a wider dispersion of OTA production at 0.90a(w)/25°C compared to 0.98a(w)/25°C, while CV at 0.98a(w)/37°C was similar to that at 0.98a(w)/25°C. In order to address the strain variability in growth initiation and prove the well-established notion of reducing OTA in foods by preventing fungal growth, a greater number of strains should be included when developing models for conditions that are suboptimal both for a(w) for OTA production and temperature levels for growth.

The aim of this study was to determine the effect of some pre-harvest fungicides on growth and aflatoxin (AF) production of three Aspergillus flavus strains found in Capsicum powder. Each isolate, previously isolated from paprika, chilli and smoked paprika, was inoculated on yeast extract sucrose agar and on a 3% paprika extract agar medium supplemented with different fungicides and incubated at 20 and 30°C during 7 days. Growth measurements were obtained on days 3, 5 and 7, and the AF production was determined on day 7. The significance of the effects of the factors (strain, medium, temperature, time and fungicides) and their interaction over colony diameter and AF production was determined. Temperature constrained the effectiveness of fungicides in reducing growth, the fungicides being most effective at 20°C. The efficacy of the fungicides over AF production depended on the medium used and temperature. The most effective fungicides in inhibiting growth and AF production, regardless of the strain tested or applied conditions, were tebuconazole 25% and mancozeb 80% applied at a concentration of 0.75 and 3.5 g l(-1), respectively. Care should thus be taken in the choice of a suitable fungicide because their effectiveness may depend on intra-specific variation and temperature. Moreover, it is necessary to take into account that the most efficient fungicide in reducing growth is not always the best choice for pre-harvest treatments because it may promote AF production. Thus, the best fungicide is the one that can simultaneous prevent growth and AF production.

The aim of this study was to determine the co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) in paprika and chilli samples purchased in Spain, using HPLC with fluorescence detection. The occurrence of mycotoxin in 64 paprika samples was 59% for AFs, 98% for OTA and 39% for ZEA, whereas in the 35 chilli samples, the contamination was 40% for AFs, 100% for OTA and 46% for ZEA. None of the samples had AFs levels higher than the legally allowable limits. Regarding the co-occurrence of mycotoxins, 75% of paprika samples and 65% of chilli samples contained more than one mycotoxin. Chilli samples generally had lower concentrations of AFB1, AFB2, total AFs and OTA than had paprika samples. The high incidence of OTA contamination suggests that additional legislation may be required to for these kinds of spices.

A survey of the fungal contamination and occurrence of aflatoxins and ochratoxin A (OTA) in 50 pistachio nut samples collected from commercial stores around Catalonia (Spain) was carried out. Aspergillus flavus was found in 30% of samples, whereas Aspergillus section Nigri, Aspergillus ochraceus, and Penicillium verrucosum were found in 40%, 2% and 26% of samples, respectively. A total of 204 fungal isolates were obtained; 70.8% of A. flavus isolates were able to produce aflatoxin B1 and B2, whereas OTA production capacity was detected in 54.4% of the A. section Nigri biseriate isolates. Aflatoxins were detected in five samples (10% of total), all exceeding the maximum legal limit set for aflatoxin B1 (2 µg kg(-1)) or for total aflatoxins (4 µg kg(-1)), with one sample having a very high level of contamination (1134.5 µg kg(-1)). Only one sample of pistachio showed OTA contamination at 0.67 µg kg(-1). Simultaneous contamination by aflatoxins and OTA was not observed in these samples. It was concluded that the frequency of aflatoxins and OTA contamination in pistachio samples available in the Spanish market is relatively low. Thus, no significant contribution to health risk should be expected from aflatoxin consumption through pistachios among healthy consumers. However, the existence of single samples with an extremely high aflatoxin concentration reinforces the need for effective prevention strategies in pistachio processing and commercialization.

The effect of three alternating temperatures cycles (20/30, 20/37 and 25/42 °C) and photoperiod on growth and Ochratoxin A (OTA) production of six isolates of Aspergillus carbonarius on synthetic nutrient medium were investigated. The different temperature regimes affected significantly both the mycelial growth and the OTA production. The best growth and OTA production were recorded at 20/30 °C. The isolates from the region of Baddar produced the highest OTA yields. A 24 h light cycle generally enhanced the growth of A. carbonarius . Growth rates cycles of 11 h/13 h light/darkness and 24 h darkness were often similar for individual isolates, such conditions enhanced OTA production in two of the six isolates tested.

Ochratoxin A (OTA) is a fungal toxic secondary metabolite that can be found in several foodstuffs and thereby ingested by humans. One way to assess exposure of humans to OTA is the determination of the levels of this mycotoxin in blood plasma from a certain population. Such studies have been done in many countries, both in healthy people and nephropathy patients. Relationships with individual characteristics were investigated in several cases. Thus, most studies found no correlation with age, either with gender. However, the few studies that found correlation between OTA plasma levels and gender showed that men presented the highest values. When sampling was done over more than one season, the highest OTA plasma levels were found mostly in summer. Differences within regions of a country were related to dietary habits of each area. OTA levels of group populations showed variations from year to year, whereas intraindividual repetitions showed no specific trend. Daily intake of the toxin can be estimated from OTA plasma concentrations by the Klaassen equation. OTA toxicokinetics are considered in this review. Calculated daily intake of OTA by different studies did not overpass the proposed tolerable daily intakes of OTA.

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.

This research was undertaken to evaluate the effects of the natural phytochemicals trans-cinnamic acid (CA) alone at concentrations of 20 and 25 mM, ferulic acid (FA) at concentration of 30 mM and two mixtures, CA-FA (20+30 mM) and CA-FA (25+30 mM) on natural maize mycoflora, Aspergillus section Flavi population and aflatoxin B1 production. These studies were carried out in maize grain in relation to a water activity of 0.99, 0.97 and 0.94. CA at 25 mM and the mixture CA-FA (25+30 mM) were the most effective treatments at inhibiting natural maize mycoflora at all aw assayed after 11 and 35 days of incubation at 25 °C. In general, 20 mM CA caused complete inhibition of Aspergillus section Flavi population at all aw values tested during all incubation period without an additional inoculum. 20 mM CA and 25 mM CA showed the major inhibitory effect on aflatoxin B1 accumulation of control and Aspergillus section Flavi additionally inoculated during all incubation periods. The data showed that CA and FA could be considered as effective fungitoxicants for natural maize mycoflora and aflatoxigenic fungi in the aw range 0.99 to 0.94. The information obtained shows promise for controlling aflatoxigenic fungi in stored maize.

BACKGROUND: The aim of this study was to screen the multicontamination by mycotoxins in a wide variety of aromatic and/or medicinal herb samples collected in Spain. Mycotoxins studied were aflatoxins (AFs), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), T-2 toxin (T-2), citrinin and fumonisins (FBs). Mycotoxins were analysed by ELISA after a clean-up step with multifunctional columns (AFs, ZEA, DON, FBs and T-2) or polyamide column (citrinin). RESULTS: Of the 84 samples analysed 99% were contaminated with T-2, 98% with ZEA, 96% with AFs, 63% with OTA, 62% with DON, 61% with citrinin and 13% with FBs. Nearly 87% of samples contained four or more mycotoxins simultaneously, being AFs, T-2 and ZEA the mycotoxins co-existing in almost every sample. 100% of the samples in our study were multicontaminated. CONCLUSION: This study shows that this kind of commodity could be an important source of mycotoxin contamination and, in general, this contamination is not limited to only one group of mycotoxins. Mycotoxin contamination on artichoke immature florets, boldus leaves, burdock leaves, dandelion plant, frangula bark, ginkgo leaves, lemon verbena leaves, olive leaves, red tea leaves, ribgrass leaves, St Mary's thistle seeds, spearmint leaves, star anise fruit, vervain and white tea leaves has been described for the first time. Finally, this is the first report on DON and T-2 presence in herbs. No study of this kind has been previously developed in Spain. Copyright © 2009 Society of Chemical Industry

BACKGROUND: Accumulation of patulin in apples and pears and subsequently in their by-products is caused mostly by Penicillium expansum which causes blue mould. Fruit pH and other parameters are sometimes characteristic of a certain variety and thus the use of particular varieties might affect patulin content in the final products. The aim of this study was to evaluate the influence of fruit variety and pH on patulin accumulation. Patulin accumulation in both apple and pear juices at different pH and also in different apple varieties (Golden, Gala and Fuji) and pear varieties (Blanquilla and Conference) was assessed. RESULTS: The pH of juices significantly influenced patulin accumulation, especially in apple juice in which the highest amounts of patulin were detected at pH 3.5. In fruits, the pH values of the substrate were determinant only under cold storage. Thus, Golden apples, which presented a lower pH, accumulated more patulin at 1 ◦ C. However, this trend was not observed at other temperatures in which varieties with higher amounts of organic acids (Golden and Fuji apples) accumulated most patulin. In the pear varieties, significant differences in pH did not lead to significant differences in patulin accumulation. CONCLUSION: Although pH influences patulin accumulation, other factors such as organic acid content may play an important role. Concerning the apple and pear varieties we studied, it seems that rather than variety, other parameters like pH or acidic content may be more important. These parameters vary significantly depending on the degree of ripeness of the fruit. Gala apple, which is used in Spanish juice production, was the only variety that accumulated dramatically higher amounts of patulin.  2008 Society of Chemical Industry

Aims: To evaluate the effect of Botrytis cinerea growth on ochratoxin A (OTA) production by Aspergillus carbonarius and degradation. Methods and Results: OTA-producing A. carbonarius and B. cinerea were grown on grape-like medium at 20°C for 7 days. Radii of colonies were daily recorded and OTA was analysed. In addition, each B. cinerea isolate was inoculated on grape-like synthetic nutrient medium (SNM) paired with each A. carbonarius isolate at a distance of 45 mm. Botrytis cinerea isolates were also grown in OTA-spiked SNM. Growth rates of B. cinerea and A. carbonarius were 20 and 7·5 mm day−1, respectively. The growth of the colonies of each species stopped when they contacted each other in paired cultures. OTA production by A. carbonarius in the contact area was affected by B. cinerea, but no clear trend was observed. All B. cinerea isolates showed to degrade between 24·2% and 26·7% of OTA from spiked SNM. Conclusions: The ecological advantage of B. cinerea, in terms of growth rate, vs. OTA-producing Aspergillus in some wine-growing regions and its ability to degrade OTA may explain the low levels of this toxin in noble wines. Significance and Impact of the Study: At determinate conditions, the presence of B. cinerea in grapes with A. carbonarius may help in reducing OTA accumulation.

Owing to their usual conditions of production in countries with warm and humid climates and to poor storage conditions, products derived from Capsicum sp. are susceptible to fungal contamination and development, which can lead to the accumulation of mycotoxins in these foods. Thus, products as chilli or paprika can be contaminated with fungal toxins, such as aflatoxins, ochratoxins and other mycotoxins, which pose a serious risk to public health. This study reviews the main aspects regarding mycotoxin contamination of Capsicum, in the context of the importance of this product in a global market and approaches the effect of processing on final contamination of foods, as well as reviews the analytical methodology commonly employed in fungal and mycotoxin analysis in these types of products.

Penicillium expansum is the most important fungal species causing spoilage in cold stored apples. P. expansum produces patulin, a toxic secondary metabolite. The aim of this work was to study the changes in growth parameters and patulin accumulation that may occur when two different isolates of P. expansum germinate and develop in the same wound. The effect of the inoculum size was also studied. For that purpose, apples were point inoculated with two different isolates of P. expansum at two different conidia concentrations. Both isolates were also inoculated together. Apples were cold stored at 1 °C or incubated at 20 °C until lesion diameter reached 2.5 cm. Patulin from decayed tissue was analysed at the end of cold storage/incubation. It seems that intraspecific interactions occur. The interactions resulted in slower growth in which patulin content was lower. A competition for resources followed by either a “combat-like” interaction may be involved in this. Higher inoculum size led to shorter lag phase, which reflects the importance of the hygiene in packinghouses facilities. The inoculum size also affected growth rate and patulin accumulation, which suggests that interactions between individuals of the same strain may occur. Competition for resources and a combat-like interaction may take place. In vitro studies should be carried out in order to assess the patterns of the interactions observed.

Growth of a range of 14 common food spoilage fungal species was evaluated along time as a function of both colony diameter and ergosterol content on malt extract agar. Growth was assessed under different environmental conditions following a central composite design. The suitability of using either linear, Gompertz's or Baranyi's models for primary modelling of the results was tested. Regarding colony diameters, using either linear or asymptotic Baranyi's function gave better estimations of growth rate and lag phase when no asymptotic trend was observed. When a decrease in growth rate was observed with time, standard Baranyi's model was chosen, although the search for new mechanistic models specific for moulds would probably improve the estimations. The use of Gompertz equation led, in general, to overestimated parameters. Ergosterol showed good performance as a fungal growth indicator for the whole range of species. Finally, significant correlation coefficients were found between ergosterol and colony diameters, suggesting that both parameters may be useful for primary modelling and thus for subsequent secondary modelling.