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Mycotoxins are secondary metabolites produced by moulds in food that are considered a substantial issue in the context of food safety, due to their acute and chronic toxic effects on animals and humans. Therefore, new accurate methods for their identification and quantification are constantly developed in order to increase the performance of extraction, improve the accuracy of identification and reduce the limit of detection. At the same time, several industrial practices have shown the ability to reduce the level of mycotoxin contamination in food. In particular, a decrease in the amount of mycotoxins could result from standard processes naturally used for food processing or by procedures strategically introduced during processing, with the specific aim of reducing the amount of mycotoxins. In this review, the current methods adopted for accurate analyses of mycotoxins in cereals (aflatoxins, ochratoxins, trichothecenes, fumonisins) are discussed. In addition, both conventional and innovative strategies adopted to obtain safer finished products from common cereals intended for human consumption will be explored and analysed. © 2018 Society of Chemical Industry.

The production of biodegradable and edible films with desired mechanical characteristics and gas barrier properties represents one of the most advanced challenges in the field of food wrapping and coating. New edible films can serve not only to provide food with physical protection but also to reduce loss of their moisture, to restrict absorption of oxygen, to lessen migration of lipids, to improve their mechanical handling features, and as materials, to apply in direct contact with internal food to realize a multilayer food packaging. Polymers derived from natural products, like carbohydrates and proteins, offer the greatest opportunities as component of edible films since their biodegradability and environmental compatibility are assured and they can also supplement the nutritional value of specific foods. However, excessive water solubility and poor water vapor barrier properties, and often poor mechanical resistance, have their application limited until the present time. Numerous studies have been carried out to improve their properties by preparing composite and multi-component films or by physically and chemically crosslinking their natural components. In the present review we summarize the main results obtained by crosslinking with the enzyme transglutaminase different proteins contained in multi-component pectin- and chitosan-based edible films, having the aim to create environmentally-friendly "bioplastics" with mechanical and permeability properties similar to the ones exhibited by plastics of petrochemical origin.

The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.

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A Ca(2+)-independent microbial TGase (transglutaminase) isolated from Streptoverticillium mobaraense was used to obtain whey protein containing novel dairy products. We evaluated the difference both in the curd formation time as well as in the hardness and deformability of the cheese obtained from cow's milk in the presence or absence of the enzyme. The results of our experiments showed that the milk coagulation time was dependent on the step in cheese manufacture at which TGase was added. We analysed the deformability and the hardness of the dairy products obtained either by adding both TGase and the milk-clotting enzyme to the milk sample at the same time or by adding TGase after treating the milk sample for 30 min with the clotting enzyme and cutting the obtained coagulum. TGase treatment conferred a strongly decreased protein content to derived whey. Moreover, when further amounts of whey were added to the milk during the manufacturing process in the presence of TGase, whey-protein-enriched dairy products could also be obtained. Our findings may lead to new biotechnologies for the re-utilization of by-products from dairy plants and contribute to reduction of environmental pollution from whey-protein disposal.

Introduction: New fungal species are increasingly reported in immunocompromised patients. Saprochaete clavata (S. clavata), an ascomycetous fungus formerly called Geotrichum clavatum, is intrinsically resistant to echinocandins and is often misidentified.Objective: We describe a cluster of seven S. clavata infections in hospitalized hematology patients who developed this rare fungemia within a span of 11 months. Three of the seven patients died. Identification of the isolates was determined only with the Saramis database of VitekMS system and sequencing of the internal transcribed spacer (ITS) region. Clonal relatedness of the isolates was determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) analysis; clonal correlation between the strains was investigated by means of phylogenetic analysis, based on single-nucleotide variants (SNPs). Clinical presentation, 1-3 β-Dglucan (BG) and galactomannan (GM) antigen results and analysis of possible sources of contamination are also described with a prospective case-control study of the outbreak.Results: MALDI-TOF MS-Vitek (bioMerieux, Marcy l'Etoile, France) failed to identify the six isolates, while SARAMIS (bioMerieux, Marcy l'Etoile, France) identified the isolates as S. clavata. Initially, Vitek 2 identified the strains as Geotrichum capitatum in two of the seven cases. Molecular identification gave 99% homology with S. clavata. BG was positive in three out of six patients (range 159 to >523 pg/ml), GM results were always negative. All the isolates were resistant to echinocandins (anidulafungin, micafungin, and caspofungin) and Fluconazole, but susceptible to Flucytosine and Voriconazole. One isolate showed acquired resistance to Flucytosine and Amphotericin Frontiers in Microbiology | www.frontiersin.org 1 January 2020 | Volume 11 | Article 84Lo Cascio et al.Saprochaete clavata Outbreak Investigation Strategy B during treatment. Both the correlation-based dendrograms obtained by MALDI-TOF MS (Bruker Daltonics) and MS-Vitek not only clustered six of the seven bloodstream infection (BSI) isolates in the same group, but also showed their strong relatedness. Phylogenetic analysis using SNPrelate showed that the seven samples recorded during the investigation period clustered together. We observed a split between one case and the remainder with a node supported by a z-score of 2.3 (p-value = 0.021) and 16 mutations unique to each branch.Conclusion: The use of proteomics for identification and evaluation of strain clonality in outbreaks of rare pathogens is a promising alternative to laborious and time-consuming molecular methods, even if molecular whole-genome sequencing (WGS) typing will still remain the reference method for rare emergent pathogens.