Fungal diversity and metabolomic profiles in GM and isogenic non-GM maize cultivars from Brazil.

There is little knowledge of the microbial diversity, mycotoxins and associated secondary metabolites in GM maize and isogenic non-GM cultivars (cvs). This study has quantified the microbial populations and dominant fungal genera in 6 cvs of each type representative of herbicide, pesticide or stacked resistance to both. The predominant mycotoxins and targeted metabolomics profiles were also compared between the two sets of cvs. This showed that the overall fungal populations were 8.8 CFUs g-1 maize. The dominant genera, isolated from maize samples, whether surface-sterilised or not, in all maize cvs were Fusarium, followed by Penicillium, Aspergillus and occasionally Cladosporium and Alternaria. The analysis of the targeted metabolomics showed that approx. 29 different metabolites were detected. These were dominated by fumonisins and minor Penicillium spp. metabolites (questiomycin A and rugulovasine A). Interestingly, the range and number of mycotoxins present in the GM cvs were significantly lower than in the non-GM maize samples. This suggests that while the fungal diversity of the two types of maize appeared to be very similar, the major contaminant mycotoxins and range of toxic secondary metabolites were much lower in the GM cvs.

Community movement and COVID-19: a global study using Google's Community Mobility Reports.

Google's 'Community Mobility Reports' (CMR) detail changes in activity and mobility occurring in response to COVID-19. They thus offer the unique opportunity to examine the relationship between mobility and disease incidence. The objective was to examine whether an association between COVID-19-confirmed case numbers and levels of mobility was apparent, and if so then to examine whether such data enhance disease modelling and prediction. CMR data for countries worldwide were cross-correlated with corresponding COVID-19-confirmed case numbers. Models were fitted to explain case numbers of each country's epidemic. Models using numerical date, contemporaneous and distributed lag CMR data were contrasted using Bayesian Information Criteria. Noticeable were negative correlations between CMR data and case incidence for prominent industrialised countries of Western Europe and the North Americas. Continent-wide examination found a negative correlation for all continents with the exception of South America. When modelling, CMR-expanded models proved superior to the model without CMR. The predictions made with the distributed lag model significantly outperformed all other models. The observed relationship between CMR data and case incidence, and its ability to enhance model quality and prediction suggests data related to community mobility could prove of use in future COVID-19 modelling.

Beyond Toxin Transport: Novel Role of ABC Transporter for Enzymatic Machinery of Cereulide NRPS Assembly Line.

Nonribosomal peptide synthetases (NRPSs) and polyketide synthetases (PKSs) play a pivotal role in the production of bioactive natural products, such as antibiotics and cytotoxins. Despite biomedical and pharmaceutical importance, the molecular mechanisms and architectures of these multimodular enzyme complexes are not fully understood. Here, we report on an ABC transporter that forms a vital part of the nonribosomal peptide biosynthetic machinery. Emetic Bacillus cereus produces the highly potent, mitochondrial active nonribosomal depsipeptide cereulide, synthesized by the NRPS Ces. The ces gene locus includes, next to the structural cesAB genes, a putative ABC transporter, designated cesCD Our study demonstrates that tethering of CesAB synthetase to the cell membrane by CesCD is critical for peptide assembly. In vivo studies revealed that CesAB colocalizes with CesCD on the cell membrane, suggesting direct involvement of this ABC transporter in the biosynthesis of a nonribosomal peptide. Mutation of cesCD, disrupting the assembly of the CesCD complex, resulted in decreased interaction with CesAB and, as a consequence, negatively affected cereulide biosynthesis. Specific domains within CesAB synthetase interacting with CesC were identified. Furthermore, we demonstrated that the structurally similar BerAB transporter from Bacillus thuringiensis complements CesCD function in cereulide biosynthesis, suggesting that the direct involvement of ABC transporter in secondary metabolite biosynthesis could be a widespread mechanism. In summary, our study revealed a novel, noncanonical function for ABC transporter, which is essential for megaenzyme functionality of NRPS. The new insights into natural product biosynthesis gained may facilitate the discovery of new metabolites with bioactive potential.IMPORTANCE This study revealed a novel, potentially conserved mechanism involved in the biosynthesis of microbial natural products, exemplified by the mitochondrial active depsipeptide cereulide. Similar to other bioactive substances, such as the last-resort antibiotics vancomycin and daptomycin, the antitumor drug cryptophycin or the cholesterol-lowering agent lovastatin, cereulide is synthesized nonribosomally by multienzyme machinery, requiring the concerted actions of multiple proteins to ensure correct product assembly. Given the importance of microbial secondary metabolites in human and veterinary medicine, it is critical to understand how these processes are orchestrated within the host cells. By revealing that tethering of a biosynthetic enzyme to the cell membrane by an ABC transporter is essential for nonribosomal peptide production, our study provides novel insights into synthesis of microbial secondary metabolites, which could contribute to isolation of novel compounds from cryptic secondary metabolite clusters or improve the yield of produced pharmaceuticals.

Online behavioural patterns for Coronavirus disease 2019 (COVID-19) in the United Kingdom.

The current coronavirus (COVID-19) pandemic offers a unique opportunity to conduct an infodemiological study examining patterns in online searching activity about a specific disease and how this relates to news media within a specific country. Google Trends quantifies volumes of online activity. The relative search volume was obtained for 'Coronavirus', 'handwashing', 'face mask' and symptom related keywords, for the United Kingdom, from the date of the first confirmed case until numbers peaked in April. The relationship between online search traffic and confirmed case numbers was examined. Search volumes varied over time; peaks appear related to events in the progression of the epidemic which were reported in the media. Search activity on 'Coronavirus' correlated well against confirmed case number as did 'face mask' and symptom-related keywords. User-generated online data sources such as Google Trends may aid disease surveillance, being more responsive to changes in disease occurrence than traditional disease reporting. The relationship between media coverage and online searching activity is rarely examined, but may be driving online behavioural patterns.

Efficacy of metabolites of a Streptomyces strain (AS1) to control growth and mycotoxin production by Penicillium verrucosum, Fusarium verticillioides and Aspergillus fumigatus in culture.

The objectives of this study were to determine the efficacy of metabolites of a Streptomyces strain AS1 on (a) spore germination, (b) mycelial growth, (c) control of mycotoxins produced by Penicillium verrucosum (ochratoxin A, OTA), Fusarium verticillioides (fumonisins, FUMs) and Aspergillus fumigatus (gliotoxin) and (d) identify the predominant metabolites involved in control. Initial screening showed that the Streptomyces AS1 strain was able to inhibit the mycelial growth of the three species at a distance, due to the release of secondary metabolites. A macroscopic screening system showed that the overall Index of Dominance against all three toxigenic fungi was inhibition at a distance. Subsequent studies showed that the metabolite mixture from the Streptomyces AS1 strain was very effective at inhibiting conidial germination of P. verrucosum, but less so against conidia of A. fumigatus and F. verticillioides. The efficacy was confirmed in studies on a conducive semi-solid YES medium in BioScreen C assays. Using the BioScreen C and the criteria of Time to Detection (TTD) at an OD = 0.1 showed good efficacy against P. verrucosum when treated with the Streptomyces AS1 extract at 0.95 and 0.99 water activity (aw) when compared to the other two species tested, indicating good efficacy. The effective dose for 50% control of growth (ED50) at 0.95 and 0.99 aw were approx. 0.005 ng/ml and 0.15 µg/ml, respectively, with the minimum inhibitory concentration (MIC) at both aw levels requiring > 40 µg/ml. In addition, OTA production was completely inhibited by 2.5 µg/ml AS1 extract at both aw levels in the in vitro assays. Ten metabolites were identified with four of these being predominant in concentrations > 2 µg/g dry weight biomass. These were identified as valinomycin, cyclo(L-Pro-L-Tyr), cyclo(L-Pro-L-Val) and brevianamide F.

Carbon dioxide production as an indicator of Aspergillus flavus colonisation and aflatoxins/cyclopiazonic acid contamination in shelled peanuts stored under different interacting abiotic factors.

Aspergillus flavus is the main xerophylic species colonising stored peanuts resulting in contamination with aflatoxins (AFs) and cyclopiazonic acid (CPA). This study evaluated the relationship between storage of shelled peanuts under interacting abiotic conditions on (a) temporal respiration (R) and cumulative CO2 production, (b) dry matter losses (DMLs) and (c) aflatoxin B1 (AFB1) and CPA accumulation. Both naturally contaminated peanuts and those inoculated with A. flavus were stored for 7-days under different water activities (aw; 0.77-0.95) and temperatures (20-35°C). There was an increase in the temporal CO2 production rates in wetter and warmer conditions, with the highest respiration at 0.95 aw + A. flavus inoculum at 30°C (2474 mg CO2kg-1h-1). The DMLs were modelled to produce contour maps of the environmental conditions resulting in maximum/minimum losses. Maximum mycotoxin contamination was always at 0.95 aw although optimal temperatures were 25-30°C for AFs and 30-35°C for CPA. These results showed a correlation between CO2 production and mycotoxin accumulation. They also provide valuable information for the creation of a database focused on the development of a post-harvest decision support system to determine the relative risks of contamination with these mycotoxins in stored shelled peanuts.

Effect of interacting conditions of water activity, temperature and incubation time on Fusarium thapsinum and Fusarium andiyazi growth and toxin production on sorghum grains.

This study examined the effect of interacting conditions of water activity (aW, 0.995, 0.98 and 0.95) and temperature (15, 25 and 30 °C) on growth rate of two Fusarium thapsinum and one F. andiyazi strains isolated from sorghum in Argentina. In addition, the effect of interacting conditions (aW × temperature × incubation time (7, 14, 21 and 28 days)) on mycotoxin production (moniliformin (MON), fusaric acid (FA) and fusarin C (FUS C)) on a sorghum grain substrate was evaluated. Statistical analysis showed that aW and temperature significantly affected growth of both species, mainly the aW. Incubation time significantly influenced mycotoxin production by both species as well, mostly for FA. Maximum growth rates of the F. thapsinum strains were obtained at the highest aW (0.995) and 25 °C and growth rate decreased as aW and temperature were reduced. The same growth profile was observed for F. andiyazi RCFA09 (maximum growth rates at 0.995-25 °C). Mycotoxin production by both species was detected at the highest aW levels whereas at 0.95 aW only low amounts of MON were produced by F. thapsinum. Maximum MON and FUS C production by both F. thapsinum strains was observed at 0.995 aW and 25-30 °C after 28 days of incubation. Also, F. thapsinum strains showed maximum FA production at the highest aW and temperature but after 14 days; after this incubation time toxin levels significantly decreased. The responses to aW and temperature of F. andiyazi were similar to that of F. thapsinum strains in relation to FA and FUS C production. Maximum levels of FA were detected at the highest aW after 14 days of incubation at 25-30 °C. Fusarin C was produced at all assayed temperatures but maximum levels were detected at 30 °C and 0.995 aW after 28 days of incubation. Two-dimensional profiles on the interactions of aW by temperature were developed from these data to identify conditions that indicate a significant risk from MON, FA and FUS C accumulation on sorghum grains. The results of this study suggest that sorghum grains could be colonized by these species and toxin production can occur, especially during development stages under field conditions at high water activity of grains or during grain storage if the drying process is slow or deficient. To our knowledge, this study described for the first time FUS C production by F. thapsinum and F. andiyazi under interacting conditions of aW, temperature and incubation time on sorghum grains.

Influence of storage environment on maize grain: CO2 production, dry matter losses and aflatoxins contamination.

Poor storage of cereals, such as maize can lead to both nutritional losses and mycotoxin contamination. The aim of this study was to examine the respiration of maize either naturally contaminated or inoculated with Aspergillus flavus to examine whether this might be an early and sensitive indicator of aflatoxin (AF) contamination and relative storability risk. We thus examined the relationship between different interacting storage environmental conditions (0.80-0.99 water activity (aw) and 15-35°C) in naturally contaminated and irradiated maize grain + A. flavus on relative respiration rates (R), dry matter losses (DMLs) and aflatoxin B1 and B2 (AFB1-B2) contamination. Temporal respiration and total CO2 production were analysed by GC-TCD, and results used to calculate the DMLs due to colonisation. AFs contamination was quantified at the end of the storage period by HPLC MS/MS. The highest respiration rates occurred at 0.95 aw and 30-35°C representing between 0.5% and 18% DMLs. Optimum AFs contamination was at the same aw at 30°C. Highest AFs contamination occurred in maize colonised only by A. flavus. A significant positive correlation between % DMLs and AFB1 contamination was obtained (r = 0.866, p < 0.001) in the irradiated maize treatments inoculated with A. flavus. In naturally contaminated maize + A. flavus inoculum loss of only 0.56% DML resulted in AFB1 contamination levels exceeding the EU legislative limits for food. This suggests that there is a very low threshold tolerance during storage of maize to minimise AFB1 contamination. This data can be used to develop models that can be effectively used in enhancing management for storage of maize to minimise risks of mycotoxin contamination.

Mycotoxin contamination of foods in Southern Africa: A 10-year review (2007-2016).

Major staple foods in Southern Africa are prone to mycotoxin contamination, posing health risks to consumers and consequent economic losses. Regional climatic zones favor the growth of one or more main mycotoxin producing fungi, Aspergillus, Fusarium and Penicillium. Aflatoxin contamination is mainly reported in maize, peanuts and their products, fumonisin contamination in maize and maize products and patulin in apple juice. Lack of awareness of occurrence and risks of mycotoxins, poor agricultural practices and undiversified diets predispose populations to dietary mycotoxin exposure. Due to a scarcity of reports in Southern Africa, reviews on mycotoxin contamination of foods in Africa have mainly focused on Central, Eastern and Western Africa. However, over the last decade, a substantial number of reports of dietary mycotoxins in South Africa have been documented, with fewer reports documented in Botswana, Lesotho, Malawi, Mozambique, Zambia and Zimbabwe. Despite the reported high dietary levels of mycotoxins, legislation for their control is absent in most countries in the region. This review presents an up-to-date documentation of the epidemiology of mycotoxins in agricultural food commodities and discusses the implications on public health, current and recommended mitigation strategies, legislation, and challenges of mycotoxin research in Southern Africa.

Putative neuromycotoxicoses in an adult male following ingestion of moldy walnuts.

A tremorgenic syndrome occurs in dogs following ingestion of moldy walnuts, and Penicillium crustosum has been implicated as the offending fungus. This is the first report of suspected moldy walnut toxicosis in man. An adult male ingested approximately eight fungal-infected walnut kernels and after 12 h experienced tremors, generalized pain, incoordination, confusion, anxiety, and diaphoresis. Following symptomatic and supportive treatment at a local hospital, the man made an uneventful recovery. A batch of walnuts (approximately 20) was submitted for mycological culturing and identification as well as for mycotoxin analysis. Penicillium crustosum Thom was the most abundant fungus present on walnut samples, often occurring as monocultures on isolation plates. Identifications were confirmed with DNA sequences. The kernels and shells of the moldy walnuts as well as P. crustosum isolates plated on yeast extract sucrose (YES) and Czapek yeast autolysate (CYA) agars and incubated in the dark at 25 °C for 7 days were screened for tremorgenic mycotoxins and known P. crustosum metabolites using a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. A relatively low penitrem A concentration of only 1.9 ng/g was detected on the walnut kernels when compared to roquefortine C concentrations of 21.7 µg/g. A similar result was obtained from P. crustosum isolates cultured on YES and CYA, with penitrem A concentrations much lower (0.6-6.4 µg per g mycelium/agar) compared to roquefortine C concentrations (172-1225 µg/g). The authors surmised that besides penitrem A, roquefortine C might also play an additive or synergistic role in intoxication of man.

Characterization of fungi in office dust: Comparing results of microbial secondary metabolites, fungal internal transcribed spacer region sequencing, viable culture and other microbial indices.

Recent developments in molecular and chemical methods have enabled the analysis of fungal DNA and secondary metabolites, often produced during fungal growth, in environmental samples. We compared 3 fungal analytical methods by analysing floor dust samples collected from an office building for fungi using viable culture, internal transcribed spacer (ITS) sequencing and secondary metabolites using liquid chromatography-tandem mass spectrometry. Of the 32 metabolites identified, 29 had a potential link to fungi with levels ranging from 0.04 (minimum for alternariol monomethylether) to 5700 ng/g (maximum for neoechinulin A). The number of fungal metabolites quantified per sample ranged from 8 to 16 (average = 13/sample). We identified 216 fungal operational taxonomic units (OTUs) with the number per sample ranging from 6 to 29 (average = 18/sample). We identified 37 fungal species using culture, and the number per sample ranged from 2 to 13 (average = 8/sample). Agreement in identification between ITS sequencing and culturing was weak (kappa = -0.12 to 0.27). The number of cultured fungal species poorly correlated with OTUs, which did not correlate with the number of metabolites. These suggest that using multiple measurement methods may provide an improved understanding of fungal exposures in indoor environments and that secondary metabolites may be considered as an additional source of exposure.

Multimycotoxin analysis of South African Aspergillus clavatus isolates.

Aspergillus clavatus poisoning is a neuromycotoxicosis of ruminants that occurs sporadically across the world after ingestion of infected feedstuffs. Although various toxic metabolites are synthesized by the fungus, it is not clear which specific or group of mycotoxins induces the syndrome. A. clavatus isolates were deposited in the culture collection of the Biosystematics Division, Plant Protection Research Institute, Agricultural Research Council during incidences of livestock poisoning (1988-2016). Six isolates were still viable and these plus three other South African isolates that were also previously deposited in the collection were positively identified as A. clavatus based on morphology and ß-tubulin sequence data. The cultures were screened for multiple mycotoxins using a liquid chromatography-tandem mass spectrometric (LC-MS/MS) method. Twelve A. clavatus metabolites were detected. The concentrations of the tremorgenic mycotoxins (i.e., tryptoquivaline A and its related metabolites deoxytryptoquivaline A and deoxynortryptoquivaline) were higher than patulin and cytochalasin E. Livestock owners should not feed A. clavatus-infected material to ruminants as all the South African A. clavatus isolates synthesized the same compounds when cultured under similar conditions.

Causal agents of Fusarium head blight of durum wheat (Triticum durum Desf.) in central Italy and their in vitro biosynthesis of secondary metabolites.

Durum wheat samples harvested in central Italy (Umbria) were analyzed to: evaluate the occurrence of the fungal community in the grains, molecularly identify the Fusarium spp. which are part of the Fusarium head blight (FHB) complex and characterize the in vitro secondary metabolite profiles of a subset of Fusarium strains. The Fusarium genus was one of the main components of the durum wheat fungal community. The FHB complex was composed of eight species: Fusarium avenaceum (61%), F. graminearum (22%), F. poae (9%), F. culmorum (4%), F. proliferatum (2%), F. sporotrichioides (1%), F. sambucinum (0.5%) and F. langsethiae (0.5%). F. graminearum population was mainly composed of the 15-acetyldeoxynivalenol chemotype, while, F. culmorum population was composed of the 3-acetyldeoxynivalenol chemotype. In vitro characterization of secondary metabolite biosynthesis was conducted for a wide spectrum of substances, showing the mycotoxigenic potential of the species complex. F. avenaceum strains were characterized by high enniantin and moniliformin production. F. graminearum strains were in prevalence deoxynivalenol producers. F. poae strains were characterized by a high biosynthesis of beauvericin like the F. sporotrichioides strain which was also found to be a high T-2/HT-2 toxins producer. Production of aurofusarin, butenolide, gibepyrone D, fusarin C, apicidin was also reported for the analyzed strains.

Newly discovered ergot alkaloids in Sorghum ergot Claviceps africana occurring for the first time in Israel.

Sorghum ergot is a disease caused commonly by C. africana. In 2015, ergot was identified for the first time in sorghum fields in Israel, leading to measures of eradication and quarantine. The aims of the study were to identify the ergot species by molecular and ergot alkaloid profile analysis, to determine the ergot alkaloid profile in pure honeydew and in infected sorghum silages and to estimate the safety of sorghum silages as a feed source. C. africana was rapidly and reliably identified by microscopical and molecular analysis. Dihydroergosine was identified as the major ergot alkaloid. Dihydrolysergol and dihydroergotamine were identified for the first time as significant ergot alkaloid components within the C. africana sclerotia, thereby providing for the first time a proof for the natural occurrence of dihydroergotamine. The sorghum silages were found to be safe for feed consumption, since the ergot alkaloids and the regulated mycotoxins were below their regulated limits.

Microbial secondary metabolites in homes in association with moisture damage and asthma.

We aimed to characterize the presence of microbial secondary metabolites in homes and their association with moisture damage, mold, and asthma development. Living room floor dust was analyzed by LC-MS/MS for 333 secondary metabolites from 93 homes of 1-year-old children. Moisture damage was present in 15 living rooms. At 6 years, 8 children had active and 15 lifetime doctor-diagnosed asthma. The median number of different metabolites per house was 17 (range 8-29) and median sum load 65 (4-865) ng/m(2) . Overall 42 different metabolites were detected. The number of metabolites present tended to be higher in homes with mold odor or moisture damage. The higher sum loads and number of metabolites with loads over 10 ng/m(2) were associated with lower prevalence of active asthma at 6 years (aOR 0.06 (95% CI <0.001-0.96) and 0.05 (<0.001-0.56), respectively). None of the individual metabolites, which presence tended (P < 0.2) to be increased by moisture damage or mold, were associated with increased risk of asthma. Microbial secondary metabolites are ubiquitously present in home floor dust. Moisture damage and mold tend to increase their numbers and amount. There was no evidence indicating that the secondary metabolites determined would explain the association between moisture damage, mold, and the development of asthma.

Quantitation of multiple mycotoxins and cyanogenic glucosides in cassava samples from Tanzania and Rwanda by an LC-MS/MS-based multi-toxin method.

A multi-mycotoxin method based on liquid chromatography/tandem mass spectrometry (LC-MS/MS) was used for a mycotoxin survey in 627 samples of processed cassava collected from different districts across Tanzania and Rwanda after the method performance for this matrix had been determined. Matrix effects as well as extraction efficiencies were found to be similar to most other previously investigated matrices with the exception of distinct matrix effects in the negative ionisation mode for early eluting compounds. Limits of detection were far below the regulatory limits set in the European Union for other types of commodities. Relative standard deviations were generally lower than 10% as determined by replicates spiked on two concentration levels. The sample-to-sample variation of the apparent recoveries was determined for 15 individually spiked samples during three different analytical sequences. The related standard deviation was found to be lower than 15% for most of the investigated compounds, thus confirming the applicability of the method for quantitative analysis. The occurrence of regulated mycotoxins was lower than 10% (with the exception of zearalenone) and the related limits were exceeded only in few samples, which suggests that cassava is a comparatively safe commodity as regards mycotoxins. The most prevalent fungal metabolites were emodin, kojic acid, beauvericin, tryptophol, 3-nitropropionic acid, equisetin, alternariol methylether, monocerin, brevianamide F, tenuazonic acid, zearalenone, chrysophanol, monilifomin, enniatins, apicidin and macrosporin. The related concentrations exceeded 1 mg kg(-1) only in few cases. However, extremely high levels of cyanogenic plant toxins, which had been previously added to the method, were observed in few samples, pointing out the need for improved post-harvest management to decrease the levels of these compounds.

Mycotoxins in corn and wheat silage in Israel.

Silage is an important feed source for intensive dairy herds worldwide. Fungal growth and mycotoxin production before and during silage storage is a well-known phenomenon, resulting in reduced nutritional value and a possible risk factor for animal health. With this in mind, a survey was conducted to determine for the first time the occurrence of mycotoxins in corn and wheat silage in Israel. A total of 30 corn and wheat silage samples were collected from many sources and analysed using a multi-mycotoxin method based on LC-MS/MS. Most mycotoxins recorded in the present study have not been reported before in Israel. Overall, 23 mycotoxins were found in corn silage; while wheat silage showed a similar pattern of mycotoxin occurrence comprising 20 mycotoxins. The most common post-harvest mycotoxins produced by the Penicillium roqueforti complex were not found in any tested samples, indicative of high-quality preparation and use of silage. Moreover, none of the European Union-regulated mycotoxins--aflatoxin B1, ochratoxin, T-2 toxin, diacetoxyscirpenol and deoxynivalenol--were found above their limits of detection (LODs). The Alternaria mycotoxins--macrosporin, tentoxin and alternariol methyl ether--were highly prevalent in both corn and wheat silage (>80%), but at low concentrations. The most prominent (>80%) Fusarium mycotoxins in corn silage were fusaric acid, fumonisins, beauvericin, monilifomin, equisetin, zearalenone and enniatins, whereas in wheat silage only beauvericin, zearalenone and enniatins occurred in more than 80% of the samples. The high prevalence and concentration of fusaric acid (mean = 765 µg kg⁻¹) in Israeli corn silage indicates that this may be the toxin of highest potential concern to dairy cow performance. However, more data from different harvest years and seasons are needed in order to establish a more precise evaluation of the mycotoxin burden in Israeli silage.

Fungal and mycotoxin assessment of dried edible mushroom in Nigeria.

In order to determine whether dried mushrooms are a foodstuff that may be less susceptible to infection by toxigenic molds and consequently to mycotoxin contamination, 34 dried market samples were analyzed. Fungal population was determined in the samples by conventional mycological techniques and molecular studies, while the spectrum of microbial metabolites including mycotoxins was analyzed by a liquid chromatography tandem mass spectrometric method covering 320 metabolites. Molds such as Fusarium, Penicillium, Trichoderma and aflatoxigenic species of Aspergillus (Aspergillus flavus and Aspergillus parvisclerotigenus) were recovered from all samples at varying levels. None of the mycotoxins addressed by regulatory limits in the EU was positively identified in the samples. However, 26 other fungal metabolites occurred at sub- to medium µg/kg levels in the samples, including aflatoxin/sterigmatocystin bio-precursors, bis-anthraquinone derivatives from Talaromyces islandicus, emerging toxins (e.g. enniatins) and other Fusarium metabolites, and clavine alkaloids. Although little is known on the toxicology of these substances, the absence of aflatoxins and other primary mycotoxins suggests that dried mushrooms may represent a relatively safe type of food in view of mycotoxin contamination.

Mycotoxins and fungal metabolites in groundnut- and maize-based snacks from Nigeria.

This exploratory study was aimed at investigating the spectrum of fungal metabolites in the processed food and snacks. Twenty types of snacks made separately from groundnut (n = 10), maize (n = 8) and a combination of groundnut and maize (n = 2) were analysed for naturally occurring mycotoxins and other fungal metabolites by a liquid chromatography-tandem mass spectrometric multi-mycotoxin method. A total of 18, 21 and 32 metabolites were detected and quantified in the groundnut-, groundnut/maize- and maize-based snacks, respectively. Aflatoxins contaminated 2, 3 and 5 of the groundnut/maize-, groundnut- and maize-based snacks at concentrations up to 14, 1041 and 74 µg kg(-1), respectively. Thus, the National Agency for Food and Drug Administration and Control (NAFDAC) recommended limit of 20 µg kg(-1) for aflatoxins was exceeded in 6 of the 20 snacks. Fumonisins contaminated all the maize- and groundnut/maize-based snacks with higher concentrations in the maize-based snacks (mean = 218.7 µg kg(-1)) compared with the groundnut/maize-based snacks (mean = 178.5 µg kg(-1)). Up to 26 different metabolites were found to co-occur in the same samples, thus posing an additional threat to the consumers due to possible additive and/or synergistic effects.