A total of 602 samples of organically and conventionally grown barley, oats and wheat was collected at grain harvest during 2002-2004 in Norway. Organic and conventional samples were comparable pairs regarding cereal species, growing site and harvest time, and were analysed for Fusarium mould and mycotoxins. Agronomic and climatic factors explained 10-30% of the variation in Fusarium species and mycotoxins. Significantly lower Fusarium infestation and concentrations of important mycotoxins were found in the organic cereals. The mycotoxins deoxynivalenol (DON) and HT-2 toxin (HT-2) constitute the main risk for human and animal health in Norwegian cereals. The impacts of various agronomic and climatic factors on DON and HT-2 as well as on their main producers F. graminearum and F. langsethiae and on total Fusarium were tested by multivariate statistics. Crop rotation with non-cereals was found to reduce all investigated characteristics significantly - mycotoxin concentrations as well as various Fusarium infestations. No use of mineral fertilisers and herbicides was also found to decrease F. graminearum, whereas lodged fields increased the occurrence of this species. No use of herbicides was also found to decrease F. langsethiae, but for this species the occurrence was lower in lodged fields. Total Fusarium infestation was decreased with no use of fungicides or mineral fertilisers, and with crop rotation, as well as by using herbicides and increased by lodged fields. Clay and to some extent silty soils seemed to reduce F. graminearum in comparison with sandy soils. Concerning climate factors, low temperature before grain harvest was found to increase DON; and high air humidity before harvest to increase HT-2. F. graminearum was negatively correlated with precipitation in July but correlated with air humidity before harvest. F. langsethiae was correlated with temperature in July. Total Fusarium increased with increasing precipitation in July. Organic cereal farmers have fewer cereal intense rotations than conventional farmers. Further, organic farmers do not apply mineral fertiliser or pesticides (fungicides, herbicides or insecticides), and have less problem with lodged fields. The study showed that these agronomic factors were related to the infestation of Fusarium species and the concentration of mycotoxins. Hence, it is reasonable to conclude that farming system (organic versus conventional) impacts Fusarium infestation, and that organic management tends to reduce Fusarium and mycotoxins. However, Fusarium infestation and mycotoxin concentrations may be influenced by a range of factors not studied here, such as local topography and more local climate, as well as cereal species and variety.

A total of 602 samples of cereals, consisting of organically and conventionally produced barley, oats and wheat, were collected at harvest during 2002-2004 in Norway. Organic and conventional cereals were sampled in comparable numbers regarding cereal species, localisation and harvest time, and analysed for Fusarium mould and mycotoxins. Fusarium infestation and mycotoxin content were dependent on cereal species and varied year-by-year. However, in all cereal species, Fusarium infestation and levels of important mycotoxins were significantly lower when grown organically than conventionally. Concerning the most toxic trichothecenes, HT-2 and T-2 toxin, lower concentrations were found in organic oats and barley. Wheat was not contaminated by HT-2 and T-2, but lower concentrations of deoxynivalenol (DON) and moniliformin (MON) were found when organically produced. For mycotoxins considered to constitute the main risk to humans and animals in Norwegian cereals, i.e. HT-2 in oats and DON in oats and wheat, the median figures (mean levels in brackets) were as follows: HT-2 in organic and conventional oats were 20 (80) and 62 (117) mu g/kg, DON in organic and conventional oats were 24 (114) and 36 (426) mu g/kg, and DON in organic and conventional wheat were 29 (86) and 51 (170) mu g/kg, respectively. Concentrations of HT-2 and T-2 in the samples were strongly correlated (r = 0.94). Other mycotoxins did not show a significant correlation to each other. Both HT-2 and T-2 concentrations were significantly correlated with infestation of F. langsethiae (r = 0.65 and r = 0.60, respectively). Concentrations of DON were significantly correlated with F. graminearum infestation (r = 0.61). Furthermore, nivalenol (NIV) was significantly correlated with infestation of F. poae (r = 0.55) and MON with F. avenaceum (r = 0.37). As lower Fusarium infestation and mycotoxin levels were found in organic cereals, factors related to agricultural practice may reduce the risk of contamination with Fusarium mycotoxins. Studies of these issues will be presented separately.

Norwegian grain samples (73 oats, 75 barley, 80 wheat) from the 2000 to 2002 growing seasons were examined for contamination with five different enniatins and the association between the found concentrations and the prevalence or infection level with several common Fusarium species investigated. Enniatin B was the fungal metabolite with the highest prevalence (100%) and the highest maximum concentration (5800 μg/kg, wheat). The maximum concentration of all five enniatins together in a single sample was 7400 μg/kg (wheat). Enniatin concentrations were correlated with several independent variables, among them grain species. Beauvericin was only sporadically detected in barley and wheat and at concentrations just above the limit of detection of 3 μg/kg, while amounts up to 120 μg/kg were found in oats. The likelihood of detecting enniatin A1 as well as the concentrations of enniatins B and B1 could be mainly related to infection with Fusarium avenaceum/arthrosporioides, and the likelihood of detecting beauvericin could be related to infection with Fusarium poae. This survey indicates that the prevalence of enniatins A1, B and B1 in Norwegian grain is high, and that enniatin B concentrations of above 1000 μg/kg are common in barley and wheat.

Partial translation elongation factor 1 alpha (TEF-1alpha) gene and intron sequences are reported from 148 isolates of 11 species of the anamorph genus Fusarium; F. avenaceum (syn. F. arthrosporioides), F. cerealis, F. culmorum, F. equiseti, F.flocciferum, F. graminearum, F. lunulosporum, F. sambucinum, F. torulosum, F. tricinctum and F. venenatum. The sequences were aligned with TEF-1alpha sequences retrieved from 35 isolates of F. kyushuense, F. langsethiae, F. poae and F. sporotrichioides in a previous study, and 39 isolates of F. cerealis, F. culmorum, F. graminearum and F. pseudograminearum retrieved from sequence databases. The 222 aligned sequences were subjected to phylogenetic analyses using maximum parsimony and Bayesian Markov Chain Monte Carlo maximum likelihood statistics. Support for internal branching topologies was examined by Bremer support, bootstrap and posterior probability analyses. The resulting trees were largely congruent. The taxon groups included in the sections Discolor, Gibbosum and Sporotrichiella sensu Wollenweber & Reinking (1935) all appeared to be polyphyletic. All species were monophyletic except F. flocciferum that was paraphyletic, and one isolate classified as F. cfr langsethiae on the basis of morphology that grouped with F. sporotrichioides. Mapping of toxin profiles, host preferences and geographic origin onto the DNA based phylogenetic tree structure indicated that in particular the toxin profiles corresponded with phylogeny, i.e. phylotoxigenic relationships were inferred. A major distinction was observed between the trichothecene and non-trichothecene producers, and the trichothecene producers were grouped into one clade of strictly type A trichothecene producers, one clade of strictly type B trichothecene producers and one clade with both type A and type B trichothecene producers. Furthermore, production of the type A trichothecenes T-2/HT-2 toxins are associated with a lineage comprising F. langsethiae and F. sporotrichioides. The ability to produce zearalenone was apparently gained parallel to the ability to produce trichothecenes, and later lost in a derived sublineage. The ability to produce enniatins is a shared feature of the entire study group, with the exception of the strict trichothecene type B producers and F. equiseti. The ability to produce moniliformin seems to be an ancestral feature of members of the genus Fusarium which seems to have been lost in the clades consisting of trichothecene/zearalenone producers. The aims of the present study were to determine the phylogenetic relationships between the different species of Fusarium commonly occurring on Norwegian cereals and some of their closest relatives, as well as to reveal underlying patterns such as the ability to produce certain mycotoxins, geographic distribution and host preferences. Implications for a better classification of Fusarium are discussed and highlighted.

The morphological variation, secondary metabolite profiles and restriction fragment length polymorphisms (RFLPs) of PCR amplified intergenic spacer (IGS) ribosomal DNA (rDNA) were studied in 27 isolates of Fusarium equiseti, 25 isolated from Norwegian cereals and 2 from soil obtained from the IBT culture collection (BioCentrum, Technical University of Denmark). All 27 isolates were tested for production of fusarochromanone (FUSCHR), zearalenone (ZEA) and the trichothecenes: 15-monoacetoxy-scirpentriol (MAS), diacetoxy-scirpenol (DAS), T-2 and HT-2 toxins, T2-triol, neosolaniol (NEO), deoxynivalenol (DON), nivalenol (NIV) and 4-acetylnivalenol (Fus-X). The trichothecenes were analysed by GC-MS in a selected ion monitoring mode, while FUSCHR was determined by ion pair HPLC with fluorometric detection and production of ZEA by TLC. For amplification of IGS rDNA primers CNL12 and CNS1 were applied. IGS rDNA was digested with the four restriction enzymes: AvaII, CfoI, EcoRI and Sau3A. In addition, we sequenced the IGS rDNA region of three of the Norwegian isolates. There were two morphological types among the Norwegian strains of F. equiseti, type I with short apical cells (dominating) and type II with long apical cells, with four haplotypes identified based on the RFLP data. Variation in secondary metabolite profiles within and between the morphological groups was observed and the levels of produced toxins were: FUSCHR 3000–42,500 and 25–30 ng/g, NIV 20–2500 and 120–700 ng/g, FUS-X 20–15,000 and 0 ng/g, DAS 30–7500 and 0–600 ng/g, and MAS 10–600 and 0–500 ng/g, for strains with short and long apical cells, respectively. NEO was detected in 16/27 strains tested (all morphotype I). All but four strains of type I (these four lacked a restriction site for EcoRI) had identical RFLP profiles. The isolates of type II had two haplotypes. The IGS sequence similarity data indicated differences between these morphotypes corresponding to two separate lineages apparently at the species level.

Norwegian grain samples (73 oats, 75 barley, 83 wheat) from the 2000–02 growing seasons were examined for contamination with moniliformin, and the association between the fungal metabolite and the number of kernels infected with common Fusaria was investigated. Before quantification of moniliformin using ion pairing reversed-phase high-performance liquid chromatography with diode array ultraviolet light detection, all samples were extracted using acetonitrile/water (84/16) and disposable strong anion exchange columns used for clean up. The limit of detection was 40 μg kg−1. Moniliformin was found in 25, 32 and 76% of the barley, oats and wheat samples, respectively. The maximum concentrations of moniliformin in barley, oats and wheat were 380, 210 and 950 μg kg−1, respectively. At the same time, the prevalence and infection level of the moniliformin-producing F. avenaceum/arthrosporioides was as high as 100 and >53% on average, respectively. Moniliformin concentrations were significantly correlated to the variables grain species, growing season and infection with F. avenaceum/arthrosporioides and F. culmorum. The survey indicates that the prevalence of moniliformin in Norwegian grain is high, especially in wheat. On the other hand, field conditions in Norway do not seem to favour contamination of grain with high levels of moniliformin.

A new species of Fusarium, Fusarium langsethiae, is described, illustrated and discussed. This species is isolated from kernels of oats, wheat and barley in several European countries. Morphologically, the species resembles Fusarium poae. It is differentiated from F. poae by slower growth, less aerial mycelium and absence of odour; its napiform or globose conidia are borne in the aerial mycelium on the agar surface on often bent phialides which exhibit sometimes more than one opening, whereas those of F. poae are produced on straight monophialides mostly in the aerial mycelium. No sporodochial conidia are formed by F. langsethiae even under near-UV light (nUV). Based on morphological characters, the species is placed in the section Sporotrichiella.

The occurrence and geographic distribution of species belonging to the genera Alternaria and Fusarium in grains of reduced and of acceptable quality were studied post-harvest in 1997 and 1998. A total of 260 grain samples of wheat, barley and oats was analysed. The distribution of Alternaria and Fusarium spp. varied significantly in samples of reduced quality compared with acceptable samples. Alternaria spp. dominated in the acceptable samples with A. infectoria group as the most frequently isolated and most abundant species group of this genus while Fusarium spp. dominated in samples of reduced quality. The most frequently isolated Fusarium spp. from all samples were F. avenaceum, F. poae, F. culmorum and F. tricinctum. Other important toxigenic Fusarium spp. isolated were F. graminearum and F. equiseti. The infection levels of F. graminearum and F. culmorum were significantly higher in the samples of reduced quality. The results indicated a negative interaction between F. graminearum and Alternaria spp. as well as between F. graminearum and other Fusarium spp.

Phylogenetic relationships between four Fusarium species were studied using parts of the nuclear translation elongation factor-1 alpha (EF-1α) gene as a phylogenetic marker. Sequences from 12 isolates of Fusarium poae, 10 isolates of Fusarium sporotrichioides and 12 isolates of Fusarium langsethiae yielded 4, 5 and 5 haplotypes, respectively. In addition, we included one isolate of Fusarium kyushuense. The aligned sequences were subjected to neighbor-joining (NJ), maximum parsimony and maximum likelihood (ML) analyses. The results from the different analyses were highly concordant. The EF-1α-based phylogenies support the classification of F. langsethiae as a separate taxon in the section Sporotrichiella of Fusarium, as the closest sister taxon to F. sporotrichioides, while F. kyushuense is the sister taxon to F. poae. This corresponds well with the ability of F. langsethiae and F. sporotrichioides to produce T-2 and HT-2 toxins. In contrast, morphological characters indicate a closer relationship between F. langsethiae and F. poae on the one hand, and between F. sporotrichioides and F. kyushuense on the other hand.

An integrated systematic study was carried out to clarify the taxonomical position and relationship of Fusarium langsethiae to other taxa within the Fusarium section Sporotrichiella. Strains of this species were compared with strains of the closely related species Fusarium poae and Fusarium sporotrichioides using a composite dataset. This set consisted of DNA sequences derived from the ribosomal internal transcribed spacer (ITS) regions, partial sequences of the ribosomal intergenic spacer (IGS) region, the β-tubulin and translation elongation factor-1 alpha (EF-1α) genes, AFLP fingerprints, chromatographic data on secondary metabolites and morphological data and growth characteristics. From these combined data, a consensus matrix was calculated by taking the mean of all pairwise distances between single isolates over all separate datasets. The consensus matrix was used as the basis for the construction of a UPGMA dendrogram and a multidimensional scaling, both of which revealed a clear separation of the three taxa. Partial IGS, EF-1α and β-tubulin sequence—as well as chromatography—and AFLP-derived similarities turned out to be comparably consistent, while ITS sequence- and morphology-derived similarity matrices were rather divergent.

In the period 1994-1996 a post-harvest survey was conducted in wheat, barley and oats to assess the occurrence and geographic distribution of Fusarium species in Norwegian cereals. The number of samples investigated was adjusted proportionally to the production of each cereal species within the regions. A total of 695 grain samples were analysed. The amount of Fusarium infection varied with cereal species and region of origin. The most frequently isolated Fusarium spp. from all samples were F. avenaceum, F. poae, F. tricinctum and F. culmorum. Other important toxigenic Fusarium spp. were F. graminearum, "powdery F. poae ", F. equiseti and F. sporotrichioides . A north-south gradient was valid for F. tricinctum, F. poae and in 1994 for "powdery F. poae ". In 1994 "powdery F. poae " was the most abundant potential producer of HT-2 and T-2 toxins in Norwegian cereals. Distribution of F. graminearum and F. culmorum demonstrated in this study , corresponded to previously reported DON-distribution, although DON seems to be produced by different species in different regions. Distribution of the isolated Fusarium species and comparison between cereals and locations are discussed.

Samples of grain, harvested in October/November of 1993 or in the spring of 1994 from fields with overwintered grain were collected. The presence of Fusarium species and the amounts of mycotoxins produced by this genus were determined, The cytotoxic properties were examined with an in vitro MTT-cell culture assay using swine kidney and VERO cells as target cells. The amount off. culmorum was significantly higher in the samples from October/November compared with the samples from spring, while the opposite was the case for F. avenaceum. Although less toxins were detected in the overwintered grain, it was more cytotoxic than the grain harvested in October/November. A significant correlation between the cytotoxicity and the amount of F. avenaceum in the samples was found.