Recent climatological research predicts a significantly wetter climate in Southern Norway as a result of global warming. Thus, the country has already experienced unusually wet summer seasons in the last three years (2010–2012). The aim of this pilot study was to apply an existing multi-analyte LC-MS/MS method for the semi-quantitative determination of 320 fungal and bacterial metabolites in Norwegian cereal grain samples from the 2011 growing season. Such knowledge could provide important information for future survey and research programmes in Norway. The method includes all regulated and well-known mycotoxins such as aflatoxins, trichothecenes, ochratoxin A, fumonisins and zearalenone. In addition, a wide range of less studied compounds are included in the method, e.g., Alternaria toxins, ergot alkaloids and other metabolites produced by fungal species within Fusarium, Penicillium and Aspergillus. Altogether, 46 metabolites, all of fungal origin, were detected in the 76 barley, oats and wheat samples. The analyses confirmed the high prevalence and relatively high concentrations of type-A and -B trichothecenes (e.g., deoxynivalenol up to 7230 µg/kg, HT-2 toxin up to 333 µg/kg). Zearalenone was also among the major mycotoxins detected (maximum concentration 1670 µg/kg). Notably, several other Fusarium metabolites such as culmorin, 2-amino-14,16-dimethyloctadecan-3-ol and avenacein Y were co-occurring. Furthermore, the most prevalent Alternaria toxin was alternariol with a maximum concentration of 449 µg/kg. A number of Penicillium and Aspergillus metabolites were also detected in the samples, e.g., sterigmatocystin in concentrations up to 20 µg/kg.

During the last ten years, Norwegian cereal grain industry has experienced large challenges due toFusarium spp. andFusarium mycotoxin contamination of small-grained cereals. To prevent severely contaminated grain lots from entering the grain supply chain, it is important to establish surveys for the most prevalentFusarium spp. and mycotoxins. The objective of our study was to quantify and calculate the associations betweenFusarium spp. and mycotoxins prevalent in oats and spring wheat. In a 6-year period from 2004-2009, 178 grain samples of spring wheat and 289 samples of oats were collected from farmers’ fields in South East Norway. The grains were analysed for 18 differentFusarium-mycotoxins by liquid chromatography – mass spectrometry. Generally, the median mycotoxin levels were higher than reported in Norwegian studies covering previous years. The DNA content ofFusarium graminearum, Fusarium culmorum, Fusarium langsethiae,Fusarium poae andFusarium avenaceum were determined by quantitative PCR. We identifiedF. graminearum as the main deoxynivalenol (DON) producer in oats and spring wheat, andF. langsethiae as the main HT-2 and T-2-toxins producer in oats. No association was observed between quantity ofF. graminearum DNA and quantity ofF. langsethiae DNA nor for their respective mycotoxins, in oats. F. avenaceum was one of the most prevalentFusarium species in both oats and spring wheat. The following ranking ofFusarium species was made based on the DNA concentrations of theFusarium spp. analysed in this survey (from high to low):F. graminearum = F. langsethiae = F. avenaceum > F. poae > F. culmorum (oats);F. graminearum = F. avenaceum > F. culmorum > F. poae = F. langsethiae (spring wheat). Our results are in agreement with recently published data indicating a shift in the relative prevalence ofFusarium species towards moreF. graminearum versusF. culmorum in Norwegian oats and spring wheat.

The increased occurrence of Fusarium-mycotoxins in Norwegian cereals over the last decade, is thought to be caused by increased inoculum resulting from more cereal residues at the soil surface as a result of reduced tillage practices. In addition, weather conditions have increasingly promoted inoculum development and infection by Fusarium species. The objective of this work was to elucidate the influence of different tillage regimes (autumn plowing; autumn harrowing; spring plowing; spring harrowing) on the inoculum potential (IP) and dispersal of Fusarium spp. in spring oats. Tillage trials were conducted at two different locations in southeast Norway from 2010 to 2012. Oat residues from the previous year’s crop were collected within a week after sowing for evaluation. IP was calculated as the percentage of residues infested with Fusarium spp. multiplied by the proportion of the soil surface covered with residues. F. avenaceum and F. graminearum were the most common Fusarium species recovered from oat residues. The IP of Fusarium spp. was significantly lower in plowed plots compared to those that were harrowed. Plowing in either the autumn or spring resulted in a low IP. Harrowing in autumn was more effective in reducing IP than the spring harrowing, and IP levels for the spring harrowed treatments were generally higher than all other tillage treatments examined. Surprisingly low levels of F. langsethiae were detected in the residues, although this species is a common pathogen of oat in Norway. The percentage of the residues infested with F. avenaceum, F. graminearum, F. culmorum and F. langsethiae generally related to the quantity of DNA of the respective Fusarium species determined using qPCR. Fusarium dispersal, quantified by quantitative PCR analysis of spore trap samples collected at and after heading, generally corresponded to IP. Fusarium dispersal was also observed to increase after rainy periods. Our findings are in line with the general understanding that plowing is a means to reduce the IP of Fusarium spp. in cereal fields. The main inoculum source for F. langsethiae remains unclear. Our results will be useful in the development of forecasting tools to calculate the risk of Fusarium in cereals.

As has been observed in several European countries, the frequency of Fusarium head blight (FHB) caused by members of the Fusarium graminearum species complex (FGSC) has increased in Norwegian cereals in recent years, resulting in elevated levels of deoxynivalenol in cereal grains. The objective of this study was to determine if this increase was associated with changes in FGSC composition within Norway. FGSC isolates collected from wheat, oats and barley in Norway during two periods, mainly 1993–1998 and 2004–2007, were characterized to determine species and trichothecene genotype composition and to assess levels of genetic variation and population structure. In vitro growth rates at different temperatures and aggressiveness in spring wheat were further characterized for a sub-selection of isolates. All Norwegian isolates were identified as F. graminearum. The 3-acetyl-deoxynivalenol (3-ADON) trichothecene type was dominant. However, isolates with the 15-ADON chemotype were detected in Norway for the first time and may represent a recent introduction of this trichothecene type. Bayesian-model based clustering and analyses of genetic differentiation indicated the persistence over the last 20 years of two sympatric and partially admixed populations of F. graminearum in Norway. Significant differences in average in vitro growth rates and aggressiveness were observed between these two populations. Our results demonstrate that the recent increase in prevalence of the FGSC in Norwegian cereals do not correspond to any dramatic changes in FGSC species or trichothecene chemotype composition. However, significant changes in population frequencies were observed among Norwegian F. graminearum.

A glutathione (GSH) adduct of the mycotoxin 4-deoxynivalenol (DON), together with a range of related conjugates, has recently been tentatively identified by LC-MS of DON-treated wheat spikelets. In this study, we prepared samples of DON conjugated at the 10- and 13-positions with GSH, Cys, CysGly, γ-GluCys and N-acetylcysteine (NAC). The mixtures of conjugates were used as standards for LC-HRMS analysis of one of the DON-treated wheat spikelet samples, as well as 19 Norwegian grain samples of spring wheat and 16 grain samples of oats that were naturally-contaminated with DON at concentrations higher than 1 mg/kg. The artificially-contaminated wheat spikelets contained conjugates of GSH, CysGly and Cys coupled at the olefinic 10-position of DON, whereas the naturally-contaminated harvest-ripe grain samples contained GSH, CysGly, Cys, and NAC coupled mainly at the 13-position on the epoxy group. The identities of the conjugates were confirmed by LC-HRMS comparison with authentic standards, oxidation to the sulfoxides with hydrogen peroxide, and examination of product-ion spectra from LC-HRMS/MS analysis. No γ-GluCys adducts of DON were detected in any of the samples. The presence of 15-O-acetyl-DON was demonstrated for the first time in Norwegian grain. The results indicate that a small but significant proportion of DON is metabolized via the GSH-conjugation pathway in plants. To our knowledge, this is the first report of in vivo conjugation of trichothecenes via their epoxy group, which has generally been viewed as unreactive. Because conjugation at the 13-position of DON and other trichothecenes has been shown to be irreversible, this type of conjugate may prove useful as a biomarker of exposure to DON and other 12,13-epoxytrichothecenes.