Report from the 1st MYCOKEY international conference Global Mycotoxin Reduction in the Food and Feed Chain held in Ghent, Belgium, 11-14 September 2017

Aflatoxins are cancer-promoting natural toxins that are produced by the fungus Aspergillus flavus and Aspergillus parasiticus. Aflatoxins have been regarded as one of the most fatal threat in food safety, due to their strong hepatotoxic, carcinogenic and teratogenic effects on human beings and animals. Among them, aflatoxin B1 (AFB1) is one of the common types which have received considerable attention. Thus, developing a rapid, simple and reliable method for determination of AFB1 in foods is very important. Herein, a preliminary study of Frster resonance energy transfer (FRET) immunoassay based on the cadmium-free quantum dots for determination of AFB1 was described. To avoid the use of hazardous heavy metals, core/shell InP/ZnS quantum dots (QDs) as an alternative for Cd-based QDs were synthesized. A silica shell with epoxy groups was used for water solubilization of the obtained nanoparticles. Then a specific anti-AFB1 monoclonal antibody (mAb) was labelled with the hydrophilic QDs via these highly reactive epoxy groups. Gel electrophoresis was used to control the binding. After that, the FRET system was developed using the Cd-free QDs conjugate as donor. Graphene oxide was selected as acceptor. In order to keep the distance between donor and acceptor close enough, the size of silica coated QDs should be controlled strictly. We found that 1-dodecantiol which was used for ligands change on the surface of InP/ZnS QDs was better than oleylamine and the optimum amount of tetraoxysilane was 5 mu L in the silylanization. Besides, only ethanol and hexane were used to wash silica coated QDs which could ensure good dispersion of QDs in water. The cut-off value for the determination of AFB1 in tube was 10 ng/mL with a preliminary study. Compared to reported FRET assays with Cd-based QDs, the developed FRET was easy-to-operate, visual and safe

Identification of toxigenic fungal species associated with maize ear rot: Calmodulin as single informative gene

Accurate identification of fungi occurring on agrofood products is the key aspect of any prevention and pest management program, offering valuable information in leading crop health and food safety. Fungal species misidentification can dramatically impact biodiversity assessment, ecological studies, management decisions, and, concerning toxigenic fungi, health risk assessment, since they can produce a wide range of toxic secondary metabolites, referred to as mycotoxins. Since each toxigenic fungal species can have its own mycotoxin profile, a correct species identification, hereby attempted with universal DNA barcoding approach, could have a key role in mycotoxins prevention strategies. Currently, identification of single marker for species resolution in fungi has not been achieved and the analysis of multiple genes is used, with the advantage of an accurate species identification and disadvantage of difficult setting up of PCR-based diagnostic assays. In the present paper, we describe our strategy to set up a DNA-based species identification of fungal species associated with maize ear rot, combining DNA barcoding approach and species-specific primers design for PCR based assays. We have (i) investigated the appropriate molecular marker for species identification, limited to mycobiota possibly occurring on maize, identifying calmodulin gene as single taxonomically informative entity; (ii) designed 17 sets of primers for rapid identification of 14 Fusarium, 10 Aspergillus, 2 Penicillium, and 2 Talaromyces species or species groups, and finally (iii) tested specificity of the 17 set of primers, in combination with 3 additional sets previously developed

Comments on "Screening and identification of novel ochratoxin A-producing fungi from grapes. Toxins 2016,8,833" - in reporting ochratoxin A production from strains of Aspergillus, Penicillium and talaromcyes

Recently a species in the genus Talaromyces, a uniseriate species of Aspergillus section Nigri and an isolate each of two widespread species, Penicillium rubens and P. commune, were reported to produce ochratoxin A. This claim was based on insufficient biological and chemical data. We propose a list of criteria that need to be met before an unexpected mycotoxin producer is reported. There have only been convincing data on ochratoxin A production for Penicillium verrucosum, P. nordicum, P. thymicola, all from Penicillium series Verrucosa, and from species in three sections of Aspergillus: section Circumdati, section Nigri and section Flavi

Editorial: Aspergillus-derived mycotoxins in the feed and food chain, volume II

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Influence of Lobesia botrana field control on black aspergilli rot and ochratoxin A contamination in grapes.

The grape berry moth Lobesia botrana is a key pest in vineyards in southern Europe. Damage caused by L. botrana larvae may encourage growth of black aspergilli, leading to ochratoxin A (OTA) accumulation in grapes. Field trials were conducted during three grape growing seasons (2005 through 2007) in Apulia, Italy, to evaluate an insecticide control strategy for L. botrana in the vineyard as an indirect method of reducing OTA contamination by reducing black aspergilli on the grapes. In the 2005 field trials, the insecticide treatment controlled attacks by L. botrana larvae and reduced OTA concentrations by up to 66% in the must samples of Negroamaro and Primitivo grape varieties. Significant differences (P < or = 0.05) also were observed in the incidence of black aspergilli. Environmental conditions in 2006 and 2007 resulted in a natural low level of infestation by L. botrana, low levels of OTA in both treated and untreated samples, and no significant differences between treated and nontreated samples. The results of our field study confirm previous reports that L. botrana is an important risk factor for OTA accumulation and are consistent with the hypothesis that controlling L. botrana in vineyards reduces OTA concentrations in grapes

Degradation of Aflatoxin B1 by a Sustainable Enzymatic Extract from Spent Mushroom Substrate of Pleurotus eryngii

Ligninolytic enzymes from white-rot fungi, such as laccase (Lac) and Mn-peroxidase (MnP), are able to degrade aflatoxin B1 (AFB1), the most harmful among the known mycotoxins. The high cost of purification of these enzymes has limited their implementation into practical technologies. Every year, tons of spent mushroom substrate (SMS) are produced as a by-product of edible mushroom cultivation, such as Pleurotus spp., and disposed at a cost for farmers. SMS may still bea source of ligninolytic enzymes useful for AFB1 degradation. The in vitro AFB1-degradative activity of an SMS crude extract (SMSE) was investigated. Results show that: (1) in SMSE, high Lac activity (4 U g−1 dry matter) and low MnP activity (0.4 U g−1 dry matter) were present; (2) after 1 d of incubation at 25 °C, the SMSE was able to degrade more than 50% of AFB1, whereas after 3 and 7 d of incubation, the percentage of degradation reached the values of 75% and 90%, respectively; (3) with increasing pH values, the degradation percentage increased, reaching 90% after 3 d at pH 8. Based on these results, SMS proved to be a suitable source of AFB1 degrading enzymes and the use of SMSE to detoxify AFB1 contaminated commodities appears conceivable

Increase of fumonisin b2 and ochratoxin a production by black Aspergillus species and oxidative stress in grape berries damaged by powdery mildew.

Powdery mildew (PM), caused by the fungus Erysiphe necator, is one of the most widespread fungal disease of grape and may cause extensive openings on the berry surface during the infection. We evaluated the effect of damage caused by PM in grape berries on the growth of and mycotoxin production by Aspergillus and on the oxidative stress in infected berries. Berries of Vitis vinifera L. cv. Negroamaro with sound skin (SS) and those naturally infected by PM were surface sterilized and inoculated with either fumonisin B2 (FB2)-producing strains of Aspergillus niger or ochratoxin A (OTA)-producing strains of Aspergillus carbonarius and incubated at 20 and 30uC. The PM berries were significantly more susceptible to both Aspergillus colonization (5 to 15 times more susceptible) and OTA and FB2 contamination (2 to 9 times more susceptible) than were SS berries. The highest toxin concentration was detected in inoculated PM berries both for OTA (9 ng/g) at 20uC and for FB2 (687 ng/g) at 30uC. In inoculated SS and PM berries, although malondialdehyde and hydrogen peroxide concentrations did not increase, the two black Aspergillus species caused a significant decrease in ascorbate content, thus inducing a pro-oxidant effect. These results indicate that grape berries affected by PM are more susceptible to black Aspergillus growth and to production and/or accumulation of FB2 and OTA. Thus, preventive control of E. necator on grape berries could reduce the mycotoxicological risk from black Aspergillus infection

Decontamination of Fumonisin B1 in maize grain by Pleurotus eryngii and antioxidant enzymes

Fumonisin B1 (FB1) is among the most common mycotoxins found in maize kernels and maize products worldwide. The microbiological process of detoxification and transformation of toxic organic pollutants is a promising method for foodstuffs decontamination. Some basidiomycetes, such as the Pleurotus eryngii species complex, include several important commercial edible varieties that can detoxify polycyclic organic compounds and a range of wastes and pollutants. We investigated the potential role of P. eryngii, one of the most consumed mushrooms, in the decontamination of FB1 in maize. In addition, selected antioxidant enzymes, (soluble peroxidase (POD), catalase (CAT) and ascorbate peroxidase), primarily involved in control of cell hydrogen peroxide levels, and lignin degradation, were analyzed, to evaluate their contributions to the molecular mechanisms of FB1 by P. eryngii. FB1 decontamination by P. eryngii and involvement of CAT and POD enzymes in the control of toxic decontamination levels of H2O2 were demonstrated. A consistent reduction of FB1 was observed at different incubation times. The average decrease levels of FB1, with respect to the control cultures, ranged from 45 to 61% (RSD < 15%). This study is a possible eco-friendly approach to reducing this mycotoxin in the feed supply chains

Occurrence of Multiple Mycotoxins in Various Types of Rice and Barley Samples in Thailand.

Contamination with multiple mycotoxins was found in rice and barley. BEA, DAS, ZEA, and aflatoxins were the mycotoxins most frequently found in samples. The assessed mycotoxin exposure does not represent a health risk for consumers