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Fusarium head blight, caused by Fusarium graminearum, continues to be one of the most important and devastating fungal diseases on cereal grains including wheat, barley, and oat crops. F. graminearum produces toxic secondary metabolites that include trichothecene type A and type B mycotoxins. There are many variants of these toxins that are produced, and in the early 2010s, a novel type A trichothecene mycotoxin known as 3ANX (7-α hydroxy,15-deacetylcalonectrin) and its deacetylated product NX (7-α hydroxy, 3,15-dideacetylcalonectrin) were identified in Minnesota, USA. In the current study, a total of 31,500 wheat spikes over a period of 6 years (2015–2020) within Manitoba, Canada, were screened for the F. graminearum pathogen, which accounted for 72.8% (2015), 98.3% (2016), 71.9% (2017), 74.4% (2018), 92.6% (2019), and 66.1% (2020) of isolations. A total of 303 F. graminearum isolates, confirmed through sequencing of the ribosomal intergenic spacer, were further investigated for variation in the gene Tri1, which was previously associated with the production of the NX toxin, as well as the accumulation of mycotoxins. A subset of these isolates, consisting of 73 isolates, which tested positive or negative for the NX-Tri1-F/R assay in this study, were cultured in vitro using rice media. Mycotoxins were quantified in these samples using mass spectrometry. Using the same rice culture, genomic DNA was isolated, and the Tri1 coding sequence along with its flanking regions (upstream and downstream of the Tri1 gene) was amplified and sequenced. Deoxynivalenol (DON) accumulated in 96% of the cultures from these isolates, while 3-acetyl deoxynivalenol (3ADON) and 3ANX mycotoxins accumulated in 66% and 63%, respectively. Nivalenol, 15-acetyl deoxynivalenol, and NX mycotoxins were detected in 62%, 36%, and 19% of samples, respectively. A significant correlation was observed between 3ADON and 3ANX (r2 = 0.87), as well as between DON and 3ANX (r2 = 0.89). This study highlights the first large identification of 3ANX- and NX-producing isolates of F. graminearum in Western Canada. In addition, it is the first identification of 15ADON chemotypes producing 3ANX in Western Canada and the first identification of 3ANX and NX-producing isolates in Manitoba, collected from wheat samples. [ABSTRACT FROM AUTHOR]

Zearalenone and radicicol are resorcylic acid lactones produced by numerous plant pathogenic fungi. Zearalenone is a non-steroidal estrogen mimic that can cause serious reproductive issues in livestock that consume contaminated feed. Radicicol is a potent inhibitor of the molecular chaperone Hsp90, which, in plants, has an important role in coordinating the host's immune response during infection. Here, we describe the identification and characterization of a soil-borne strain of the Gram-positive bacterium Aeromicrobium sp. capable of hydrolyzing the macrolide ring of resorcylic acid lactones, including zearalenone and radicicol. Proteomic analysis of biochemically enriched fractions from the isolated and cultured bacterium identified an α/β-hydrolase responsible for this activity. A recombinantly expressed and purified form of the hydrolase (termed RALH) was active against both zearalenone and radicicol. Interpretation of high-resolution mass spectrometry and NMR data confirmed the structures of the enzymatic products as the previously reported non-toxic metabolite hydrolyzed zearalenone and hydrolyzed radicicol. Hydrolyzed radicicol was demonstrated to no longer inhibit the ATPase activity of the Saccharomyces cerevisiae Hsp90 homolog in vitro. Enzymatic degradation of resorcylic acid lactones will enable insight into their biological functions. [ABSTRACT FROM AUTHOR]

Mycotoxin emergence and co-occurrence trends in Canadian grains are dynamic and evolving in response to changing weather patterns within each growing season. The mycotoxins deoxynivalenol and zearalenone are the dominant mycotoxins detected in grains grown in Eastern Canada. Two potential emerging mycotoxins of concern are sterigmatocystin, produced by Aspergillus versicolor, and diacetoxyscirpenol, a type A trichothecene produced by a number of Fusarium species. In response to a call from the 83rd Joint Expert Committee on Food Additives and Contaminants, we conducted a comprehensive survey of samples from cereal production areas in Ontario, Canada. Some 159 wheat and 160 corn samples were collected from farms over a three-year period. Samples were extracted and analyzed by LC-MS/MS for 33 mycotoxins and secondary metabolites. Ergosterol was analyzed as an estimate of the overall fungal biomass in the samples. In wheat, the ratio of DON to its glucoside, deoxynivalenol-3-glucoside (DON-3G), exhibited high variability, likely attributable to differences among cultivars. In corn, the ratio was more consistent across the samples. Sterigmatocystin was detected in some wheat that had higher concentrations of ergosterol. Diacetoxyscirpenol was not detected in either corn or wheat over the three years, demonstrating a low risk to Ontario grain. Overall, there was some change to the mycotoxin profiles over the three years for wheat and corn. Ongoing surveys are required to reassess trends and ensure the safety of the food value chain, especially for emerging mycotoxins. [ABSTRACT FROM AUTHOR]

This study investigated two outbreaks of spontaneous poisoning by Baccharis coridifolia (Asteraceae) in early-weaned beef calves in Tacuarembó, Uruguay. A total of 34 affected calves showed signs of salivation, anorexia, apathy, marked dehydration, and diarrhea. Deaths occurred 36–72 h after consumption and mortality varied from 37.5% to 43.3% for outbreak 1 and outbreak 2, respectively. The main pathological findings include diffuse severe necrosis of the prestomachs and lymphoid tissues. Ultrastructurally, epithelial cells of the rumen showed swelling, lysis of the organelles, degradation of intercellular attachments, and degradation of the nuclear chromatin. Using LC-MS with diagnostic fragmentation filtering, 56 macrocyclic trichothecenes including glycosyl and malonyl conjugates were identified. The total concentration of macrocyclic trichothecenes, including conjugates, was estimated to be 1.2 ± 0.1 mg/g plant material. This is the first report of these malonyl–glucose conjugates from Baccharis coridifolia. [ABSTRACT FROM AUTHOR]

The in vivo headspace extraction of volatile organic compounds from Micro-Tom tomato flowers was investigated using multiple solid phase microextraction (SPME) fibers of different properties to maximize the extraction selectivity for a nontargeted analysis. The three fibers used in this work were polydimethylsiloxane (PDMS), PDMS/divinylbenzene (DVB), and carboxen (CAR)/PDMS. Two sources for tomato flowers were used: Micro-Tom wild type (WT) and transgenic Micro-Tom overexpressing the carotenoid cleavage deoxygenase 1 gene. Gas chromatography-mass spectrometry (GC-MS) results demonstrated that the largest amounts of volatile organic compounds (VOCs) were observed with the PDMS/DVB fiber for both wild type and transgenic plants, but the CAR/PDMS and PDMS fibers contributed to the detection of selective compounds. Data revealed the presence of 45 VOCs from transgenic plants and 35 from the wild type when all three fibers were used together. Of the totalVOCs identified, 30 were common to both types of plants, but 15 were specific to the transgenic and 5 to the wild type plants. The compounds identified from Micro-Tom flowers were mainly monocyclic and bicyclic monoterpenes and sesquiterpenes, with one alkyl benzene compound. The bicyclic monoterpenes, (1R)-α-pinene, (1S)-α-pinene, and β-pinene, were found to be the most abundant molecules present in both wild type and transgenic plants. The overall advantage of maximizing the discovery of VOCs based on the selectivity differences with three SPME fibers was evident. Such a benefit is important in the nontargeted analysis of transgenic plants for detecting the production of unexpected compounds. Key words: headspace, gas chromatography-mass spectrometry (GC-MS), transgenic, volatile, insecticide. L'extraction in vivo espace de tete de composes organiques volatils (COVs) provenant de fleurs de tomates a ete etudiee en utilisant plusieurs (microextraction en phase solide, SPME) ayant des proprietes differentes afin de maximiser la selectivite d'extraction pour une analyse non ciblee. Les trois fibres utilisees dans ce travail sont le polydimethylsiloxane (PDMS), le PDMS/divinylbenzene (DVB) et le carboxen (CAR)/PDMS. Deux sources pour les fleurs de tomates ont ete utilisees: Micro-Tom de type sauvage (WT) et transgenique, sur-exprimant le gene CCD1. Les resultats de GC-MS ont demontre que les plus grandes quantites de COVs ont ete observees avec les fibres de DVB/PDMS pour les plantes de type sauvage comme pour les plantes transgeniques, mais les fibres RCA/PDMS et PDMS ont contribue a la detection de composes selectifs. Les donnees ont revele la presence de 45 COV dans les plantes transgeniques et 35 dans les plantes de type sauvage lorsque les trois fibres ont ete utilisees ensemble. Sur le total des composes organiques volatils identifies, 30 sont communs aux deux types de plantes, mais 15 etaient specifiques aux plantes transgeniques et 5 aux plantes de type sauvage. Les composes identifies a partir de fleurs Micro-Tom etaient essentiellement des monoterpenes et des sesquiterpenes, et un compose de benzene d'alkyle. Les monoterpenes, (1R)-α-pinene, α-pinene et le β-pinene etaient les molecules les plus abondantes a la fois dans les plantes de type sauvage et les plantes transgeniques. L'avantage de la maximisation de la decouverte de composes organiques volatils a partir de differences de selectivite avec trois fibres SPME etait evident. Un tel avantage est important dans l'analyse non ciblee de plantes transgeniques pour la detection de composes inattendus. Mots-cles: espace de tete, GC-MS, transgenique, volatil, insecticide., Introduction Plants interact with the environment by releasing volatile organic compounds (VOCs) into the atmosphere. Plant volatiles are produced by several organs of the plant with the majority released by [...]

Aflatoxin B1 is a potent human carcinogen produced by several species of Aspergillus mainly found on nuts and maize. Exposures in parts of Africa, Latin America and Asia can be at multiples, sometimes orders of magnitude above tolerable daily levels. Although human exposure to aflatoxin can be estimated by analysis of the diet, only determination of the serum albumin aflatoxin adduct provides a health-relevant exposure measure. The lack of a reference serum limits interlaboratory method validation and data comparisons. In this study, we synthetically produced AFB1-dialdehyde and covalently coupled it to serum albumin in human serum. This synthetic produced aflatoxin-serum reference material was used in conjunction with isotopically labelled internal standards to evaluate sample digestion methods. This showed using sufficient Pronase in the digestion step was critical to ensure complete proteolytic digestion, which occurs within 4 h. Increasing the digestion temperature from 37 °C to 50 °C also provided a benefit to the overall analysis. In addition, the use of dried blood spots and Volumetric Absorptive Microsampling (VAMS) were investigated showing samples stored with VAMS produced equivalent results to serum samples. [ABSTRACT FROM AUTHOR]