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Fungi of the genus Sepedonium have been studied for their properties as antagonist agents of various plant pathogens, as well as antibiotic and pigment producers. We investigated the antimicrobial ability of S. chrysospermum strain 704 isolated from fruit bodies of wild basidiomycetes and found to be not pathogenic on cultivated mushrooms. A strong antagonistic activity versus different pathogenic fungi and a high level of production of enzymes and antibiotic compounds were observed. The best growth conditions for the production of specific hydrolytic enzymes were determined. From fungal culture filtrate we purified and characterised a metabolite that strongly inhibit spore germination and hyphal growth of the necrotrophic pathogen Botrytis cinerea. The metabolite was identified as a 3,6-dimethyl--resorcylaldehyde, and showed a strong inhibition of the pathogen at concentrations starting from 0.05 ppm. Furthermore, a low level of phytotoxicity of the purified antibiotic, analysed by infiltration in bean leaves, was observed.

Compositional variation was examined across 13 microgreens species/subspecies representing Brassicaceae, Chenopodiaceae, Lamiaceae, Malvaceae and Apiaceae, grown in controlled environment. Macro-mineral concentrations were determined by ion chromatography, chlorophyll and ascorbate concentrations, and hydrophilic/lipophilic antioxidant potentials by spectrophotometry, and major carotenoids by HPLC-DAD. Nitrate hyper-accumulators and wide genotypic differences in Na, K and S concentrations were identified. Antioxidant capacity was highest in brassicaceous microgreens and significant genotypic variation was demonstrated in chlorophyll and carotenoid concentrations. High phenolic content was confirmed in Lamiaceae microgreens, with significant varietal differences, and alternative phenolics-rich microgreens from the Apiaceae were identified. Twenty-eight phenolic compounds were variably detected and quantitated through Orbitrap LC–MS/MS with flavonol glycosides, flavones and flavone glycosides, and hydroxycinnamic acids representing 67.6, 24.8 and 7.6% of the mean total phenolic content across species, respectively. The obtained information is critical for selecting new species/varieties of microgreens that may satisfy demand for both taste and health.

Tobacco cultivation is characterized by high amounts of waste biomasses whose disposal frequently represents a complex and expensive problem. A study was conducted to evaluate thepotential of pre-harvest light air-cured (Burley) and dark fire-cured (Kentucky) tobacco waste biomasses as a source of bioactive compounds (nutraceutical ingredients) such as polyphenols. Pre-harvest waste materials (topping fresh materials and residual stalks at final harvest) were collected to determine dry matter, total polyphenols content (TPC, Folin assay), and DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,20-azino-bis(3-ethylbenzothiazoline-60-sulfonic acid) diammonium salt) antioxidant capacity. Polyphenols quali-quantitative profiles obtained by Orbitrap Q Exactive of both tobacco types were also determined. Total pre-harvest waste biomass amounted to 3956.9 and 1304.4 kg d.w. ha−1 in light air-cured (Burley) and dark fire-cured (Kentucky) tobacco types, respectively. Polyphenols content, expressed as g kg−1 dry weight (d.w.), ranged between 4.6 and 15.7 g kg−1 d.w. and was generally greater in leaves than in stalks. Considering both leaves and stalks, the light air-cured (Burley) tobacco crop yielded 22.1 kg ha−1 of polyphenols, while the dark fire-cured (Kentucky) tobacco yielded 12.0 kg ha−1. DPPH and ABTS were significantly greater in leaves than in stalks waste biomass in both types of tobacco. The most abundant components were quinic and chlorogenic acids, rutin, and luteolin rutinoside.

In the Rural Development Plan (2014&ndash, 2020), the European Commission encouraged the conversion and supported the maintenance of organic farming. Organic olive oil (bioEVOO) production involves the use of environmentally sustainable fertilizers and the recycling of olive pomace (Pom) and olive vegetation waters (VW) to reduce the environmental impact of these wastes. An ecofriendly way to recycle olive wastes is to reuse them to extract bioactive compounds. In this study, the total phenolic compounds content, their profile and dosage, the antioxidant action in oil, pomace, and vegetation water was evaluated when the Trichoderma harzianum M10 was used as a biostimulant in agriculture. Two spectrophotometric tests (2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2&prime, azinobis (3-ethylbenzothiazoline-6-sulfonic) acid (ABTS)) evaluated the antioxidant potential of samples, a spectrophotometric method estimated total phenolic content, and an Ultra-High-Performance Liquid Chromatography (UHPLC)&ndash, Orbitrap method evaluated the phenolics profile. Our results showed that the biostimulation improved the antioxidant potential and the total concentration of phenolics in the bioEVOO and bio-pomace (bioPom) samples and mainly enhanced, among all classes of phenolic compounds, the production of the flavonoids and the secoiridoids. Moreover, they demonstrated the Trichoderma action in the mevalonate pathway to produce phenols for the first time. The decisive action of the Thricoderma on the production of phenolic compounds increases the economic value of the waste materials as a source of bioactive compounds useful for the pharmaceutical, cosmetic, and food industries.

The health advantages of extra-virgin olive oil (EVOO) are ascribed mainly to the antioxidant ability of the phenolic compounds. Secoiridoids, hydroxytyrosol, tyrosol, phenolic acid, and flavones, are the main nutraceutical substances of EVOO. Applications of beneficial microbes and/or their metabolites impact the plant metabolome. In this study the effects of application of selected Trichoderma strains or their effectors (secondary metabolites) on the phenolic compounds content and antioxidant potential of the EVOOs have been evaluated. For this purpose, Trichoderma virens (strain GV41) and Trichoderma harzianum (strain T22), well-known biocontrol agents, and two their metabolites harzianic acid (HA) and 6-pentyl-&alpha, pyrone (6PP) were been used to treat plants of Olea europaea var. Leccino and var. Carolea. Then the nutraceutical potential of EVOO was evaluated. Total phenolic content was estimated by Folin&ndash, Ciocalteau&rsquo, s assay, metabolic profile by High-Resolution Mass spectroscopy (HRMS-Orbitrap), and antioxidant activity by DPPH and ABTS assays. Our results showed that in the cultivation of the olive tree, T22 and its metabolites improve the nutraceutical value of the EVOOs modulating the phenolic profile and improving antioxidants activity.

Globe artichoke (Cynara cardunculus var. scolymus L. Fiori) is a traditional component of the Mediterranean diet, it is considered a rich source of phenolic compounds. Globe artichoke production is traditionally based on vegetatively propagated varietal types. As for pathologic and economic disadvantages of the former multiplication method, Italian artichoke breeding programs have produced some potential seed propagated hybrids. An experiment was conducted to assess the yield, mineral and polyphenolic profiles of two potential seed-propagated F1 hybrid 'Istar' and 'Romolo'. The artichoke yield recorded on 'Romolo' was significantly higher by 33% in comparison to 'Istar'. The N and K concentrations were significantly lower by 25.5% and higher by 10.1%, respectively, in 'Romolo' in comparison to 'Istar'. Among the macronutrients studied, K was the mineral with the higher concentration, with a mean of 24.9 g kg-1 dw. In addition, the Na concentration and Na/K ratio observed in both cultivars were low. Amongst the caffeoylquinic acids, 5-O-caffeoylquinic acid and 1,5-diO-caffeoylquinic acid were the main compounds. The remaining caffeoylquinic acids (i.e., 1- and 3-O-caffeoylquinic acids, 3,4-, 3,5- and 4,5-di-O-caffeoylquinic acids) were minor compounds in the analyzed samples. Regarding the cultivar effect, the 5-O-caffeoylquinic acid, apigenin 7-O-glucoronide, total caffeoylquinic acids, total apigenin derivatives, were significantly higher in 'Romolo' by 23.1, 180.0, 20.5, and 149.5%, respectively, in comparison to 'Istar', suggesting that specific cultivar should be selected to obtain the desired profile of bioactive compounds. Overall, the seed-propagated cultivars can represent a suitable alternative to the vegetative propagated ones, thus opening new perspectives for the artichoke production.

Enniatins (Enns) are mycotoxins produced by Fusarium spp. and are widely distributed contaminants of cereals and derivate products. Among the different identified enniatins, Enn B is the most relevant analogue in cereals in Europe. Therefore, the aim of this study was to investigate for the first time the occurrence of Enn B and Enn B phase I metabolites in 300 human urine samples throughout an ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Three different sample preparation procedures were evaluated and salting-out liquid-liquid extraction showed satisfactory validation results. Enn B was quantified in 83.7% of samples ranging from 0.006 to 0.391 ng/mL (average content: 0.016 ng/mL). In line with the in vitro observations with human liver microsomes, in the here analyzed samples the Enn B monooxygenated, N-demethylated and dioxygenated metabolites were tentatively found in 87.7%, 96.3% and 6.7% of samples. The data of this pilot biomonitoring survey indicate a frequent intake of enniatins in Italy, supporting further toxicological studies to provide better basis for understanding their potential effects in humans.

Enterotoxigenic Escherichia coli (ETEC) is the main bacterial cause of dehydrating infant diarrhoea in less-developed countries. Labile toxin (LT) is the major virulent factor of ETEC. Easy diagnostic tests are necessary to reduce the number of cases. Immunological methods have some drawbacks and also have important limitations. For that reason, a Liquid Chromatography coupled to UV detector technique (LC-UV) has been optimize to a rapid identification and quantification of LT from bacteria cultures. It is also important to know optimal conditions for LT and with this purpose several enterotoxigenic E. coli strains have been studied to determine the influence of glucose concentration and different culture media on LT production. LC-UV technique demonstrated to be a good method for LT quantification showing values of 15 ng/mL and 45 ng/mL for limits of detection and quantification respectively. LT quantification revealed that toxin production is directly related to the concentration of glucose added in the broth medium. Tryptic Soy Broth is the most efficient culture medium for E. coli growth and enterotoxin production.

Commercial coffee is available as a mixture of two varieties of coffee beans, namely Arabica, which is more expensive, and Canephora (Robusta), less expensive. To assess the correspondence between the composition indicated on the label and the real composition of commercially available coffee, it would be desirable to be able to differentiate between the two varieties. This would also help to avoid any possible commercial frauds. This work identifies parameters based on the fatty acid composition to differentiate between Arabica and Canephora coffee in a mixture. Total monounsaturated fatty acids (SMUFA), linolenic acid (cis18:3n–3) concentration, the 18:0/cis18:1n–9 ratio, and the SMUFA/SSFA ratio could be used to determine the relative amounts of Arabica and Canephora in a coffee blend.

Beauvericin (BEA) is a cyclic depsipeptide with antibiotic and insecticidal effects. It was discovered for the first time from the fungus Beauveria bassiana , but more significantly, is produced by several Fusarium strains, and considered a contaminant of several cereals like corn, wheat and barley. This study investigated the degradation of BEA by intracellular raw enzymes of four strains of Saccharomyces cerevisiae , named LO9, YE5, A34, and A17. The BEA at 25 mg/kg in a model solution and in corn flour was co-incubated with the raw enzymes from the four yeast strains, respectively. The reduction of BEA was evaluated using liquid chromatography coupled to a diode array detector (LC–DAD); the products formed during the co-incubation were determined by liquid chromatography coupled to mass spectrometry-linear ion trap (LC–MS-LIT). In model solution BEA reduction ranged from 83 to 100%. In corn flour treated with the intracellular raw enzymes, the BEA degradation was from 66 to 91%. A product resulted from the BEA degradation was identified.

Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains and known to have various biological activities. This study investigated the degradation of the minor Fusarium mycotoxin BEA present in the concentration of 5 mg/kg in a model solution and in different crispy breads produced with different flours typologies (corn, hole, wheat, durum wheat, soy and rice) during the heat treatment carried out in an oven at three different temperatures of 160, 180 and 200 °C and at 3, 6, 10, 15 and 20 min incubation. The concentration of the bioactive compound studied, analyzed with the technique of the liquid chromatography tandem mass spectrometry (LC–MS/MS), decreased in the experiment carried out in the model solution from 2.89 ± 0.13 mg/kg of the assay at 160 °C for 3 min until the complete degradation at 200 °C during 20 min incubation. In the experiments carried out using the crispy breads prepared with different kind of flours, as system to simulate a food preparation, the percentage of BEA degradation, resulted variable from 20 to 90%, with no a significative differences showed in the use of the different flour matrices. Also a metabolite of the thermical degradation of the mycotoxins BEA was identified using the LC–MS in the full scan mode.

Measurements of local environmental conditions, intensity of Fusarium head blight (FHB) in wheat spikes, biomass of Fusarium graminearum, F. culmorum, and F. poae (pathogens causing FHB) and concentration of the mycotoxins deoxynivalenol (DON) and nivalenol (NIV) in harvested wheat grain were obtained in a total of 150 location-years, originating in three European countries (Hungary, Ireland, United Kingdom) from 2001 to 2004. Through window-pane methodology, the length and starting time of temporal windows where the environmental variables were significantly associated with the biological variables were identified. Window lengths of 5 to 30 days were evaluated, with starting times from 18 days before anthesis to harvest. Associations were quantified with nonparametric Spearman correlation coefficients. All biological variables were significantly associated with at least one evaluated environmental variable (P ≤ 0.05). Moisture-related variables (e.g., average relative humidity, hours of relative humidity above 80%) had the highest positive correlations with the biological variables, but there also was a significant negative correlation between average temperature and several biological variables. When significant correlations were found, they were generally for all window lengths, but for a limited number of window start times (generally before anthesis for disease index and after anthesis for the toxins and late-season fungal biomasses). Semi-partial Spearman correlation coefficients were used to evaluate the relationship between the environmental variables and the concentration of DON and NIV after the effects of FHB intensity and fungal biomass on the mycotoxins were removed. Significant semi-partial correlations were found between relative humidity variables and DON, and between temperature and relative humidity variables and NIV for time windows that started after anthesis (and not for any earlier time windows). Results confirm that the environment influences disease, fungal biomass, and mycotoxin production, and help refine the time windows where the association is greatest. However, variability in the relationships was high, indicating that no single environmental variable is sufficient for prediction of disease or mycotoxin contamination.

Fumonisins are mycotoxins produced by Fusarium verticillioides that commonly contaminate maize and maize products. The present work shows the results of a comparative study of three different fermentation's techniques (solid and liquid medium of corn and a solid agarized medium) for the production of fumonisins B-1, B-2 and B-3 with strains of F. verticillioides. The solid medium of corn was the most effective in the production of fumonisins, being Fumonisin B-1 the one produced with higher concentration, so the extract obtained by solid fermentation process was used for FB1 purification. Fumonisins characterization and quantification were performed with reversed-phase high-performance liquid chromatography with electrospray ionization triple quadrupole tandem mass spectrometry. The role of production of reactive oxygen species (ROS) in Fumonisin B1 mediated toxicology has not been fully addresses in studies exploring FB1 toxicity. It is evaluated the level of ROS production in kidney cell line (VERO) exposed to 1, 5 and 10 mu M of FB1 for 0.5-100 min. The ROS level was detected using a fluorescence probe, 2',7'-dichlorofluorescein diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS. Significant increase of ROS products was observed in VERO cells at 10 mu M dose. These results indicate that ROS production by FBI on renal cells is a mechanism of fumonisin mediated toxicity. (C) 2010 Elsevier Ltd. All rights reserved.

To produce enniatin (ENs), Fusarium tricinctum CECT 20150 was grown in a liquid medium of potato (PDB), being mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semipreparative column using a mobile phase of acetonitrile/water using gradient condition. The purity of the ENs fractions was verified by analytical HPLC and LC/MS-MS. The pure fractions of ENs were utilized to study the biological activity on several mycotoxigenic moulds as Fusarium verticilloides, Fusarium sporotrichioides, Fusarium tricinctum, Fusarium poae, Fusarium oxysporum, Fusarium proliferatum, Beauveria bassiana, Trichoderma harzianum, Aspergillus flavus, Aspergillus parasiticus, Aspergillus fumigatus, Aspergillus ochraceus and Penicillium expansum. The results obtained demonstrated that in several antibiograms, ENs induced the inhibition of the grown microorganisms tested.

Enniatins (ENs) are antibiotic compounds of hexadepsipeptidic structure produced by several strains of Fusarium spp. The ENs A, A(1), B, B-1 were purified from extracts of Fusarium tricinctum grown on a solid medium of corn, by a low pressure liquid chromatography (LPLC) on reverse phase of Amberlite XAD-7 followed by semipreparative LC. The purity and the structure of the isolated compounds were confirmed by LC-MS/MS. The technique of the purification of the fungal extract enabled complete separation of the ENs A, A(1), B, B-1 with a mean purity of 97% for all the compounds. The cytoxicity of the ENs was tested in the cell lines of human origin (epithelial colorectal adenocarcinoma cells, Caco-2) by MTT assays. Only EN A(1) and B-1 evoked toxicity at the tested concentrations. The inhibitory concentration (IC50) for EN A(1) on Caco-2 cells was 12.3 mu M, whereas the IC50 produced by the EN B-1 was 19.5 mu M. This study indicates that ENs, fungal metabolites that are commonly found in corn and in general in product composed by corn, may have a toxic potential for human health. (C) 2010 Elsevier Ltd. All rights reserved.

Aspergillus niger is a significant component of the fungal community on grapes. The mycotoxin fumonisin B2 (FB2) was recently detected in grape must and wine as well as in cultures of some A. niger strains isolated from grapes and raisins. This study examined 48 strains of Aspergillus section Nigri for the presence of the fumonisin biosynthetic gene fum8 in relation to FB2 production. The fum8 gene was detected in only 11 A. niger strains, 9 of which also produced FB2. Maximum parsimony analysis based on the calmodulin gene sequence indicated that the presence/absence of fum8 is not correlated with the phylogenetic relationship of the isolates. This is the first report correlating the presence of a fumonisin biosynthetic gene with fumonisin production in A. niger from an important food crop. The results suggest that the absence of FB2 production in grape isolates of A. niger can result from the absence of at least one gene essential for production.

The use of bacillus subtilis for screening fusaric acid production by fusarium spp is described. Results allowed to set up a simple and rapid metod for fusaric acid screening in isolates.

Fusarium subglutinans (teleomorph Gibberella subglutinans, member of the Gibberella fujikuroi complex) is an important toxigenic pathogen of maize. Recently, two cryptic species (Groups 1 and 2) have been described within F. subglutinans, but little is known about the occurrence of the two groups in North America or their relative capacities to produce mycotoxins. In this study, 58 F. subglutinans strains from kernels of maize grown in Iowa, USA, were evaluated for cryptic speciation and production of the mycotoxins fusaproliferin and beauvericin. Fifty-six of the 58 strains (97%) belonged to Group 2, and two strains belonged to Group 1, based on restricted fragment length polymorphisms derived from amplification of histone H3 and beta-tubulin gene fragments. Fifty-four Group 2 strains and both Group 1 strains produced fusaproliferin at concentrations ranging from 12 to 3000 microg g(-1) of solid maize culture. None of the F. subglutinans strains from Iowa produced beauvericin at detectable amounts, although most F. subglutinans strains from Europe and elsewhere are beauvericin producers. These results indicate that F. subglutinans strains infecting maize kernels in Iowa belong almost exclusively to Group 2 and that they have a high potential for fusaproliferin production; furthermore, the results confirm an association between Group 2 genotypes and lack of beauvericin production. This is the first report characterizing the phylogenetic groups of F. subglutinans occurring in Iowa; the predominance of Group 2 suggests that populations of the fungus in Iowa and Europe remain isolated from each other. Fusaproliferin contamination of grain appears to be a risk wherever F. subglutinans occurs, but beauvericin contamination from F. subglutinans is associated only with Group 1.

Aspergillus niger has been recently found to produce fumonisin B(2) (FB(2)). Thirty-one strains belonging to four Aspergillus species isolated from grape were evaluated for FB(2) production on agar plates. Four out of eight strains of A. niger produced FB(2) (29-293 microg g(-1)). None of the strains of A. uvarum (n = 7), A. tubingensis (8) and A. carbonarius (8) produced detectable amounts of toxin. The capability to produce FB(2) was also confirmed by some A. niger strains artificially inoculated on grape berries. Natural occurrence of FB(2), at levels of 0.01 and 0.4 microg ml(-1), was found in two samples of must collected in Apulian cellars in 2007. This is the first report of FB(2) contamination in must. These findings suggest that there is a potential risk of exposure to FB(2) in the grape-wine chain for consumers and that A. niger may represent the major fumonisin-producing species among black Aspergilli occurring on grapes.

Twenty-seven preharvest maize ears affected by Fusarium poae rot (disease score 36-100%) were selected in 1998 and 1999 in Poland and examined for the occurrence of toxic hexadepsipeptides: beauvericin (BEA), enniatin A, enniatin B and enniatin B 1 The identification of F. poae was confirmed by sequence analysis of variable internal transcribed spacer regions and compared with NCBI gene bank DNA sequences. Chemical analyses were performed by HPLC-MS. In 27 ears infected by F. poae were detected: BEA (trace to 46 μg/g) in 18 samples, enniatin A (trace to 37 μg/g) in nine samples, enniatin B (trace to 47 μg/g) in 15 samples and enniatin B, (trace to 25 μg/g) in 12 samples. When 20 strains of F. poae isolated from these samples were cultured on rice, all produced BEA (1.9-75 μg/g), three enniatin A (1.8-2μg/g), 12 enniatin B (1.1-5.1 μg/g) and eight enniatin B 1 (1.2-5.2 μg/g). Occurrence and quantification of enniatin A, enniatin B and enniatin B 1 and their co-occurrence with BEA in maize kernels is reported for the first time.

Several epidemiological studies show that fruits, vegetables and cereals can play a nutraceutical role for their content of many antioxidant phytochemicals such as carotenoids, ascorbic acid and phenolics. A commercial inoculant (MICOSAT F ® ) containing arbuscular mycorrhizal fungi (AMF) could improve the nutritional value in crops. The goal of this work was to evaluate the effect of AMF on the production level of carotenoids, AsA, phenols including antocyanins and saponins, proteins, total antioxidant activity and nitrates in fruits, vegetables, legumes and durum wheat var. grecale, whose consumption is largely recommended according to Mediterranean diet. The treatment increased the antioxidant activity in strawberries (37.50%), in giant lentils (29.17%) and in durum wheat (63.63%) but decreased it in kiwi (31.81%) and in grape (19.81%). Nitrate levels decreased significantly in strawberries (39.78%) and in tomato intended for transformation (37.79%). The application of MICOSAT F ® enhanced the levels of several secondary metabolites. However, the amount of phytochemicals and respective by-products were reduced in some cases. Environmental conditions and modality of AMF inoculation could module both primary and secondary metabolites.

Ochratoxin A (OTA) is a mycotoxin produced by fungi belonging to the Penicillium and Aspergillus genera. The natural occurrence of OTA in 18 pure cocoa powder samples marketed in Italy and purchased from supermarkets, organic product shops and retail shops was investigated. Nine samples of the eighteen products had no detectable OTA contamination (below 10 ng/kg). Nine positive samples had a toxin concentration ranging between 0.22 and 0.77 μg/kg with a mean of 0.43 μg/kg. All the organic samples analyzed were negative for OTA; of 14 conventional samples analyzed, four samples (28.56%) were above the suggested legal limit (0.5 μg/kg) because they were contaminated with 0.77, 0.53, 0.67 and 0.62 μg/kg, respectively. The optimization of the extraction protocol and comparison of different brands of immunoaffinity column clean-up were carried out. The incidence (50% of examined samples) of the OTA level in Italian cocoa marketed products is an important signal of attention for consumers, because cocoa is widely used in the production of child snack foods and it contributes to OTA daily intake.

There is a urgent need to develop a rational strategy for managing Fusarium ear blight in order to reduce current reliance on routine fungicide applications, based on an objective assessment of disease risks. One of important components for such a management strategy is a fast, easy, accurate and reliable method for disease assessment. The relationship between incidence of Fusarium ear blight ear infection and number of spikelets infected on an ear (or incidence of spikelet infection) were investigated during three seasons and in four countries in order to derive a simple relationship for predicting disease at the spikelet level using ear incidence. More than half of the data sets of the number of infected spikelets on an ear could not be fitted satisfactorily by a Poisson distribution. Three two-parameter discrete distributions (negative binominal, Neyman type A and Polya-Aeppli) provided a significantly better fit than the Poisson distribution, indicating a degree of aggregation of number of infected spikelets on an ear. Taylor’s power-law satisfactorily described the observed variance–mean relationship for the number of infected spikelets on an ear; this relationship was generally consistent over years and countries. A robust relationship between incidence of ear infection and average number of infected spikelets per ear was obtained assuming a fixed variance– mean relationship and a negative binomial distribution for the number of infected spikelets. A relationship between incidences of spikelet and ear infection was also obtained based on the complementary log–log or logit transformation of ear and spikelet infection incidence. These models appeared to be consistent over years and countries and thus may be used in making practical disease management decisions involving fungicide applications.

The contamination levels of 16 different Fusarium- and Aspergillus-mycotoxins were chemically determined from randomly selected organic and conventional grain-based products purchased from Finnish and Italian markets. The cytotoxicity of the samples was analyzed with an in vitro test using feline fetal lung cells. Overall, the concentrations of the mycotoxins studied were low in all of the samples. Enniatins B and B1 as well as deoxynivalenol were the most predominant mycotoxins in the samples, being present in 97%, 97%, and 90% of the samples, respectively. The geographical origin or the agricultural practice had no influence on the mycotoxin concentrations of the samples. The babyfoods included in the samples had significantly lower concentrations of mycotoxins than the other products with a mean total mycotoxin content of 47 microg/kg compared with 99 microg/kg for the other kinds of food. All the samples evoked toxicity in the in vitro test, but no correlation between cytotoxicity and the mycotoxin concentrations was observed.

A rapid simple and economical method was described for the determination of ochratoxin A produced by Aspergillus ochraceus ITEM 5117 grown in a biofermenter in submerged culture. The ochratoxin A was determinate with RP-HPLC-FLD (reverse phase high performance liquid chromatography) with direct injection in the HPLC apparatus using a C18 column. The mycotoxin was completely resolved by using the mixture acetonitrile, water and acetic acid (49:49:2 v/v) as the mobile phase with a flow rate of 1.0 mL/min. Mean recoveries of ochratoxin A ranged from 95.36% to 103.15%. The limit of detection for ochratoxin A in medium was found to be 1 μg L −1 .

One strain of Fusarium subglutinans (ITEM-1434) isolated from maize ear rot in Poland was tested for the ability to synthesize moniliformin (MON), beauvericin (BEA) and fusaproliferin (FP) on six cereal substrates (wheat, rye, barley, oat, maize and rice kernels) for 3 weeks at 25 degrees C and on rice at three different temperatures (20, 25 and 30 degrees C). Most MON (497 micrograms/g) was produced on rice; most BEA (704 micrograms/g) on wheat or rice, and most FP (422 micrograms/g) on rye. When cultured on rice, F. subglutinans produced the highest levels of BEA and FP at 20-25 degrees C, while MON production was best at 30 degrees C.

Fusaproliferin (FP), a toxic metabolite of the world-wide maize pathogens Fusarium proliferatum and Fusarium subglutinans, was recently found to be a natural contaminant of maize. Its toxic activity on haematopoietic human cell lines and its teratogenic effects on chicken embryos has been recently proved. Therefore a sensitive, rapid, and inexpensive screening test to detect FP in agricultural commodities is necessary to protect human health. FP-hemiglutarate conjugated to modified bovine serum albumin was synthesized, characterized, and used as an antigen for raising polyclonal antibodies by immunizing rabbits. Indirect and competitive ELISA and immunoblotting analyses were performed to determine antibody specificity towards the mycotoxin. The determination of 10 µg of free FP/mL was achieved using antibodies purified by means of affinity chromatography on a FP-lysine-Sepharose column. This unsatisfactory detection limit is due to high background values; thus, this method is not competitive with traditional UV-HPLC methods.Key words: fusaproliferin, ELISA, mycotoxin, immunoassay, corn, Fusarium.

The antioxidant activity of two virgin olive oils, obtained from the same olive (Olea europaea sativa) batch but processed with different hammer crushing conditions, was evaluated by measuring their protective action towards linoleic acid peroxidation in a micellar system. The antioxidant efficiency (AE) of the oil phenolic fraction was higher when the olives were processed with a higher hammer crusher rotation rate. HPLC analysis demonstrated that one of the main derivatives of oleuropein, indicated as 3,4-DHPEA-EDA (the dialdehydic form of elenolic acid linked with 3,4- dihydroxyphenylethanol), is present only in the oil with higher AE. 3,4-DHPEA-EDA showed the greatest antioxidant ability among virgin olive oil phenols. Its greater efficiency in the micellar system in comparison with 3,4-dihydroxyphenylethanol (3,4-DHPEA) is related to its greater lipophilicity. It is suggested that the behaviour in the water-micellar environment is representative of that in a real system such as tomato-based sauce with added virgin olive oil. # 1999 Society of Chemical Industry

The aptitude of twentySaccharomyces sensu stricto strains to remove ochratoxin A from a synthetic medium containing 1.1 ng/mL, about half of the European Community limit, was evaluated using four to six mg of biomass (wet weight)/mL. Seven satins show high levels of ochratoxin A removal, 0.72-1.10 ng/mL, equivalent to 66–100% of the available toxin, and unitary removing ativity of 14.31–27.24 pg/mg of biomass. Further research will be carried out to study the mechanism of OTA removal and to confirm the ability of the most efficacious strains ofSaccharomyces sensu stricto to remove OTA from contaminated wort and grape must during alcoholic fermentation.

Aims: The aim of this study was to analyse the transcriptional regulation of enniatins (ENs) production in Fusarium avenaceum. Methods and Results: We develop a new method to quantify ENs in FDM agar medium. We performed an LC/MS/MS analysis to evaluate enniatin A, A1, B, B1 and B4 production by seven F. avenaceum strains and, in a time-course experiment, by ITEM 3404 to analyse the transcriptional regulation of the esyn1 gene. The expression profile, achieved by Real time reverse transcriptase assay, showed an activation of gene transcription at the seventh day of incubation, corresponding to the higher increase of total ENs production. Enniatin B was the most abundant ENs analogues, representing the 90% of total ENs. The relative percentage of ENs remained unaltered during the experiment. Conclusions: We reported a transcriptional regulation of esyn1 responsible for the modulation of ENs biosynthesis. Significance and Impact of the Study: Enniatins are cyclic depsipeptides metabolites with a wide range of biological activities. They are also widespread contaminants in grains and cereals due to infection by enniatin-producing Fusarium species. This is the first article describing the transcriptional regulation of esyn1 gene that modulates ENs production in Fusarium avenaceum and provides new knowledge about the molecular mechanism underlying the biosynthesis of these important fungal metabolites in this toxigenic fungal species. © 2013 The Society for Applied Microbiology.

Aflatoxins family includes a great number of lipophilic molecules produced by aerobic microscopic fungi belonging to the genus Aspergillus. The chapter describes their chemical structure, chemical and physical properties, and aspects related to their presence in food and commodities. Aflatoxins presence in food is considered a real and severe risk to consumers for their toxicity. Aflatoxins levels and frequency of foods natural contamination as reported in the scientific literature are briefly analyzed. Focus is given to the different foodstuffs that may be at risk of contamination by Aspergillus and the subsequent accumulation of aflatoxins in the food chain. Bioavailability and bioaccessibility of aflatoxins will be discussed considering that these unwanted molecules can be assumed by the humans with the diet. Bioaccessibility, that deals with the fraction of micro-nutrients released from the food matrix during digestion and gastro-intestinal available for absorption, will be discussed with reference to aflatoxins bioaccessibility of during the digestion process, considering the relationships between the food matrix and its influences on aflatoxins fate. Bioavailability of the aflatoxins assumed from the diet depends on their stability during digestion, since they are released from the food matrix (bioaccessibility) and on the efficiency of their passage through the gastro-intestinal mucosa. The term bioavailability includes the concepts of availability to the absorption, metabolism, distribution of nutrients to tissues and bioactivity and indicates the fraction of micro-nutrients absorbed by the body and the speed with which these molecules are absorbed and made available at their site of action. Despite of the practical difficulties in measuring the distribution and bioactivity of aflatoxins on a specific human body organ, the bioavailability is the fraction of an oral dose of a compound or precursor of an active metabolite that reaches the bloodstream. Bioaccessibility includes the entire sequence of events that take place during the digestion of food material that can be assimilated by the body through the epithelial cells of the gastro-intestinal mucosa. Aflatoxins are often present in very small amounts or in traces and, for this reason, a part of the chapter addresses the advanced new chromatographic and spectrometric methods described in the literature and applied to research, that can reveal, even in trace amounts, aflatoxins in biological fluids as free form or as by-products, e.g. non-covalent adducts.

Enniatins (ENs) are secondary fungal metabolites with hexadepsipeptidic chemical structure and they possess a number of potent biological activities that can contaminate several kind of food and foodstuffs increasing the exposure risk for consumers. ENs are produced by several Fusariun strains including Fusarium subglutinans, Fusarium proliferatum and Fusarium tricinctum. Production of a mixture of ENs was performed by culturing F. tricinctum ITEM 9496 on white corn as substrate. The solid culture components were dried and subsequently extracted with water/methanol (50/50 v/v, 0.5% NaCl), homogenised, filtered, extracted by ethyl acetate and analysed by liquid chromatography with diode array detection (LC-DAD). The crude extract was first separated by low pressure liquid chromatography (LPLC) and then further purified by liquid chromatography (LC), resulting in six compounds with a purity higher than 95% as calculated by 1H NMR, and with a yield of 30-300 mg per compound. The chemical structures of the ENs were determined by liquid chromatography coupled to mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR). The biological activity of the resulting ENs was determined using a mitochondrial respiration test. We discovered that all the ENs studied induced an increase in the mitochondrial respiration resulting in uncoupling of the oxidative phosphorylation. This effect was most likely due to flux of K+ ions into the mitochondrial matrix. The order of potency of the ENs derivatives was: A1 > B1 > B > A > B4 > J 1. These results suggest a correlation between the chemical structures and bioactivity and confirm the severe risks for human associated with consumption of enniatins.

Fusarium head blight (FHB) of wheat, caused by several Fusarium species, is a damaging disease, resulting not only in yield reduction but also accumulation of mycotoxins in grain. Epidemiology and management of FHB has been extensively studied worldwide. Data on FHB development and accumulation of mycotoxins were obtained in four European countries during 2001–2004 to study the effect of FHB development and environmental conditions on accumulation of deoxynivalenol (DON). The occurrence of DON was highly correlated with presence of one or more toxigenic Fusarium species. Hourly weather data recorded at each sampling site were summarised over several periods of different lengths (5–30 days) during the anthesis and pre-harvest period. All-subsets regression was used to determine the extent to which the probability of DON occurrence is related to weather variables and also the consistency of such a toxin-weather relationship. Combined with a re-sampling technique, all-subsets regression analysis showed the difficulties in identifying a single ‘best’ model of relating the probability of toxin ≥90 μg kg−1 to weather predictors. A wide range of inter-related weather predictors based on time windows around anthesis and pre-harvest were selected in different models. There were many alternative models based on weather predictors only with similar predictive power because of high correlation among weather predictors. The performance of these alternative models was generally poor, particularly in terms of the high proportion of false positive predictions (specificity was only around 0.60–0.65). Inclusion of the number of toxigenic Fusarium species at harvest into models did not improve the model sensitivity (ca. 0.75–0.80) but appreciably improved the specificity (ca. 0.70–0.75). On balance, weather summarised over a 15-day window frame led to models with better predictions than other three window frames (5, 10 and 30 days).

Patulin is a mycotoxin that contaminates pome fruits and derived products worldwide. Basidiomycete yeasts belonging to the subphylum Pucciniomycotina have been identified to have the ability to degrade this molecule efficiently and have been explored through different approaches to understand this degradation process. In this study, Sporobolomyces sp. strain IAM 13481 was found to be able to degrade patulin to form two different breakdown products, desoxypatulinic acid and ( Z ) - ascladiol. To gain insight into the genetic basis of tolerance and degradation of patulin, more than 3,000 transfer DNA (T-DNA) insertional mutants were generated in strain IAM 13481 and screened for the inability to degrade patulin using a bioassay based on the sensitivity of Escherichia coli to patulin. Thirteen mutants showing reduced growth in the presence of patulin were isolated and further characterized. Genes disrupted in patulin-sensitive mutants included homologs of Saccharomyces cerevisiae YCK2 , PAC2 , DAL5 , and VPS8 . The patulin-sensitive mutants also exhibited hypersensitivity to reactive oxygen species as well as genotoxic and cell wall-destabilizing agents, suggesting that the inactivated genes are essential for tolerating and overcoming the initial toxicity of patulin. These results support a model whereby patulin degradation occurs through a multistep process that includes an initial tolerance to patulin that utilizes processes common to other external stresses, followed by two separate pathways for degradation.

The interaction between the mycotoxin beauvericin (BEA) and 9 yeast strains of Saccharomyces cerevisiae named LO9, YE-2, YE5, YE-6, YE-4, A34, A17, A42 and A08 was studied. The biological degradations were carried out under aerobic conditions in the liquid medium of Potato Dextrose Broth (PDB) at 25 °C for 48 h and in a food/feed system composed of corn flour at 37 °C for 3 days, respectively. BEA present in fermented medium and corn flour was determined using liquid chromatography coupled to the mass spectrometry detector in tandem (LC–MS/MS) and the BEA degradation products produced during the fermentations were determined using the technique of the liquid chromatography coupled to a linear ion trap (LIT). Results showed that the S . cerevisiae strains reduced meanly the concentration of the BEA present in PDB by 86.2% and in a food system by 71.1%. All the S . cerevisiae strains used in this study showed a significant BEA reduction during the fermentation process employed.

Fusarium subglutinans is an important pathogen of maize and other commodities worldwide. We examined MRC-115 and 71 other F. subglutinans strains from various geographic areas for their ability to synthesize fusaproliferin, a novel toxic sesterterpene recently isolated from F. proliferatum. Fusaproliferin production ranged from 30 to 1,500 micrograms/g of dried ground substrate, with 33 strains producing more than 500 micrograms/g. In particular, strain MRC-115 produced as much as 1,100 to 1,300 micrograms/g. In toxicity studies of two invertebrate models, fusaproliferin was toxic to Artemia salina (50% lethal dose, 53.4 microM) and to the lepidopteran cell line SF-9 (50% cytotoxic concentration, approximately 70 microM, after a 48-h exposure). Fusaproliferin was also toxic to the human nonneoplastic B-lymphocyte cell line IARC/LCL 171 (50% cytotoxic concentration, approximately 55 microM in culture in stationary phase after a 48-h exposure). Experiments performed will cells exposed at seeding suggested a possible cytostatic effect at subtoxic concentrations.

In the present study, the occurrence of eighteen mycotoxins, nine trichothecenes (deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, neosolaniol, diacetoxyscirpenol, fusarenon-X, T-2 toxin and HT-2 toxin), three zearalenones (zearalenone, α-zearalenol and β-zearalenol), and six emergent mycotoxins, beauvericin and five enniatins (A, A1, B, B1 and B4), was monitored in different Italian organic cereals and cereal products by using a liquid chromatography coupled to triple quadrupole mass spectrometry method. A total of 93 organic cereal samples (wheat, barley, rye and oat) were collected from Italy. Limits of quantification ranged from 5 to 15 μg/kg. 80% of analyzed samples contained mycotoxins. The occurrence was 33%, 6.5%, 2%, 27%, 7%, 10% and 43% for deoxynivalenol, HT-2, T-2, nivalenol, zearalenone, beauvericin and enniatins, respectively. The major mycotoxin found was enniatin B4; it was detected in 40% of all analyzed samples and its levels ranged from 5.7 to 284.2 μg/kg. Risk assessment was evaluated by EDI calculations which were lower than TDI for all legislated Fusarium mycotoxins.

The possibilities of characterizing the heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) by liquid chromatography electrospray mass spectrometry (LC/ESI-MS) and matrix-assisted laser desorption with time-of-flight mass spectrometry (MALDI-TOF-MS) were investigated. The B subunit from recombinant E. coli (expression in Pichia pastoris) can be detected by LC/ESI-MS expressed in P. pastoris and the charge envelope signals can be observed; LC/ESI-MS and MALDI-TOF-MS analysis allowed the acquisition of labile toxin subunit B (LTB) molecular weight and preliminary structural characterization of LTB toxin. MALDI-TOF analysis after reduction and alkylation of the protein evidenced the presence of one disulfide bond in the structure of the protein. Confirmatory analysis was carried out by detection of most of the tryptic fragments of the B subunit by MALDI-TOF-MS, obtaining total coverage of the protein sequence. Possible biovariations in the toxin can mostly be determined by sequencing, where an increase of molecular mass in the N-terminal side of the protein was identified. This modification may be due to an O-GlcNAc-1-phosphorylation.

The interaction between the minor Fusarium mycotoxins BEA and 13 bacterial strains characteristic of the gastrointestinal tract as Bifidobacterium longum , Bifidobacterium bifidum , Bifidobacterium breve , Bifidobacterium adolescentes , Lactobacillus rhamnosus , Lactobacillus casei-casei , Lactobacillus plantarum , Eubacterium crispatus , Salmonella fecalis , Salmonella termofilus , Lactobacillus ruminis , Lactobacillus casei and Lactobacillus animalis was studied. The fermentations were carried out in the liquid medium of MRS during 4, 12, 16, 24 and 48 h at 37 °C, under anaerobic conditions. Levels of BEA in the fermentation liquid, on the cell walls and on the internal part of the cells were determined using liquid chromatography coupled to the mass spectrometry detector (LC-MS/MS). Results showed that the bacteria reduced the concentration of the BEA present in the medium, part of the mycotoxin was adsorbed by cell wall and part internalized by the bacteria. All the bacteria analyzed in this study showed a significant BEA reduction during the fermentation process, in particular the mean diminution resulted variable from 66 to the 83%.

Fusarium equiseti and Fusarium acuminatum are toxigenic species that contaminate cereal crops from diverse climatic regions. They are common in Spanish cereals. The information available on their phylogenetics and toxigenic profiles is, however, insufficient to assist risk evaluation. In this work, phylogenetic analyses were performed using partial sequences of the translation elongation factor gene (EF-1a) of F. equiseti and F. acuminatum strains isolated from barley and wheat from Spain and other countries. The Northern and Southern European F. equiseti strains largely separated into two phylogenetically distinct clusters. This suggests the existence of two distinct populations within this species, explaining its presence in these regions of markedly different climate. Production of type A and B trichothecenes by the Spanish strains, examined in wheat cultures using a multitoxin analytical method, indicated that F. equiseti could produce deoxynivalenol and nivalenol and other trichothecenes, at concentrations that might represent a significant risk of toxin contamination for Southern European cereals. F. acuminatum showed low intraspecific genetic variability and 58% of the strains could produce deoxynivalenol at low level. Neither species was found to produce T-2 or HT-2 toxins. The present results provide important phylogenetic and toxigenic information essential for the accurate prediction of toxigenic risk.

Finger millet (Eleusine coracana) is a subsistence crop grown in Sub-Saharan Africa and the Indian Sub-continent. Fusarium species occurring on this crop have not been reported. Approximately 13% of the Fusarium isolates recovered from finger millet growing at three different locations in eastern Uganda belong to Fusarium verticillioides, and could produce up to 18,600 µg/g of total fumonisins when cultured under laboratory conditions. These strains are all genetically unique, based on AFLP analyses, and form fertile perithecia when crossed with the standard mating type tester strains for this species. All but one of the strains is female-fertile and mating-type segregates 13:20 Mat-1:Mat-2. Three new sequences of the gene encoding translation elongation factor 1-α were found within the population. These results indicate a potential health risk for infants who consume finger millet gruel as a weaning food, and are consistent with the hypothesis that F. verticillioides originated in Africa and not in the Americas, despite its widespread association with maize grown almost anywhere worldwide.

Fumonisins are mycotoxins with cancer-promoting activity and are associated with a number of animal and human diseases. The potential risk of contamination by fumonisin B2 (FB2), although at low levels, has been demonstrated in must and wine. Black aspergilli in general and Aspergillus niger in particular are considered to be the major responsible agents of FB2 contamination in grape and its by-products. Contamination by FB2 therefore is yet another safety concern of grape and wine producers, as ochratoxin A, produced mainly by A. carbonarius, may prove to be a major mycotoxicological problem in the grape–wine chain.

We tested the compatibility of the biocontrol yeasts (Rhodosporidium kratochvilovae LS11 and Cryptococcus laurentii LS28) with the recently developed fungicides boscalid (BOSC), cyprodinil (CYPR) and fenhexamid (FENH) to create an efficient integrated approach to control blue mould on apples. The fungicide thiabendazole (TBZ), which is presently allowed for postharvest treatment of pome fruit in different countries, was also used as the control. Both the biocontrol agents (BCAs) LS11 and LS28 were compatible in vitro with BOSC and CYPR, whereas they were strongly inhibited by FENH. TBZ was compatible with LS28, while it strongly inhibited LS11. In vitro assays with some isolates of Penicillium expansum showed that the majority were resistant to TBZ, whereas they were all markedly inhibited by BOSC and CYPR. Experiments of integrated control were performed on wounded apples kept at 21 °C up to 7 days. After 4 days of storage, the combination of a low BCA concentration (5 × 106 cfu mL−1) with a low dose (25% of the label dose) of commercial formulates of BOSC or CYPR, resulted in an efficient reduction of blue mould incidence (83–100% less infection with respect to the control). Conversely, the combination of BCAs with TBZ was less effective (not more than 60% of rot reduction). When applied alone at low dosage, LS11, LS28, BOSC, CYPR and TBZ reduced Penicillium rot by 35%, 52%, 67%, 72% and 0%, respectively. After 7 days of storage, only the integrated treatment based on BCAs with BOSC or CYPR resulted in a significant rot reduction (as much as 98%). Treatments based on the utilization of the BCA LS28 or low dosage of CYPR alone were much less effective (10% and 28% rot reduction, respectively), whereas both BCAs integrated with TBZ were ineffective. Furthermore, integrated treatments (BCAs + BOSC or CYPR) resulted in lower fungicide residues and patulin (PAT) contamination in apples. Our data show that the integration of biocontrol yeasts with a low rate of the recently commercialized fungicides BOSC or CYPR could be an effective and safer strategy to control P. expansum and keep fungicide residues as well as PAT contamination in apples low.

Se describe en este artículo el descubrimiento de la degradación de la micotoxina patulina por una levadura, The infection of stored apples by the fungus Penicillium expansum causes the contamination of fruits and fruit-derived products with the mycotoxin patulin, which is a major issue in food safety. Fungal attack can be prevented by beneficial microorganisms, so-called biocontrol agents. Previous time-course thin layer chromatography analyses showed that the aerobic incubation of patulin with the biocontrol yeast Rhodosporidium kratochvilovae strain LS11 leads to the disappearance of the mycotoxin spot and the parallel emergence of two new spots, one of which disappears over time. In this work, we analyzed the biodegradation of patulin effected by LS11 through HPLC. The more stable of the two compounds was purified and characterized by nuclear magnetic resonance as desoxypatulinic acid, whose formation was also quantitated in patulin degradation experiments. After R. kratochvilovae LS11 had been incubated in the presence of 13C-labeled patulin, label was traced to desoxypatulinic acid, thus proving that this compound derives from the metabolization of patulin by the yeast. Desoxypatulinic acid was much less toxic than patulin to human lymphocytes and, in contrast to patulin, did not react in vitro with the thiol-bearing tripeptide glutathione. The lower toxicity of desoxypatulinic acid is proposed to be a consequence of the hydrolysis of the lactone ring and the loss of functional groups that react with thiol groups. The formation of desoxypatulinic acid from patulin represents a novel biodegradation pathway that is also a detoxification process.

This paper describes the first map, albeit partial, of toxigenic fungi re-isolated from grape berries collected in three out of the six most important Slovakia winemaking areas in two different periods of the harvest year 2008. Low temperatures and high relative humidity during July 2008 favoured the development of grape fungal diseases that cause rots such as Plasmopara, Uncinula, Botrytis, Metasphaeria, Elsinoe, and Saccharomycetes. In the analysed samples, the following genera of toxigenic fungi were identified in the range of 1-4%: Aspergillus, Alternaria, Cladosporium, Epicoccum, Fusarium, Penicillium, Rhizopus, Ulocladium, and Trichoderma Trichothecium, while the genera Aspergillus, Alternaria, Fusarium, and Penicillium were in the range 11-29%. A. niger, A. carbonarius, some strains of A. carbonarius-with 'crystals' and strains of A. uvarum-uniseriate were identified; these species are considered ochratoxigenic (able to produce variable amounts of toxins). In addition, a non-ochratoxigenic strain of A. ibericus and a Fusarium strain able to biosynthesize small amount of fumonisins, beauvericin, and enniatins were identified. P. expansum, able to produce citrinin, represents 29.7%, of the Penicillium genus together with P. verrucosum, P. glabrum, P. citrinum, and P. crustosum. An analysis for the identification and quantification of the main toxins: ochratoxin A, fumonisins, beauvericin, enniatins, and fusaproliferin was performed on grape samples; it was consistent with the results of the mycological analysis. Toxigenic fungi should be checked throughout the years and their occurrence compared with all environmental factors to avoid health risks.

The importance and widespread incidence of Fusarium poae as a natural contaminant of wheat in different climatic areas warrants investigation into the genetic diversity and toxin profile of a northern Italy population. Eighty-one strains of F. poae isolated from durum wheat kernels, identified by species-specific polymerase chain reaction and translation elongation factor-1alpha gene sequence analysis, were genetically characterized by the amplified fragment length polymorphism (AFLP) technique and analysed by high-pressure liquid chromatography for their ability to produce the beauvericin (BEA) and trichothecene mycotoxins. A high level of variability was observed by using AFLP analyses, with the lowest level of genetic similarity among the strains being approximately 61%. Most of the strains, 95%, produced BEA at

Carotenoids and flavonoids are the main tomatoes antioxidants, having an important role for human health. This study investigates the effects of different water regime and of the industrial processing on the concentration of these compounds in tomato fruits and in tomato products. Two biotypes of Corbarini small tomatoes were cultivated in the Sarno valley (Salerno. Italy) using three different water regimes. A biochemical characterization of the fresh and of the corresponding canned products was performed. Results show that water regime influenced the antioxidant profile of tomato fruits, with marked differences between the two biotypes. Data obtained highlight that water regime markedly influenced the productivity and the quality of the tomatoes. Results also demonstrated that industrial process increased carotenoids content without causing a significant flavonoids degradation. (C) 2010 Elsevier B.V. All rights reserved.

To obtain the bioactive compound beauvericin (BEA), Fusarium proliferatum CECT 20569 was grown on a solid medium of wheat, utilizing the technique of the solid state fermentation (SSF), being this mycotoxin purified by high performance liquid chromatography (HPLC) with a reverse phase semi-preparative column using as the mobile phase acetonitrile/water in gradient condition. The purity of the BEA was verified by analytical HPLC and liquid chromatography tandem mass spectrometry (LC/MS-MS). The pure fractions of BEA were utilized to determinate the antibiotic effects on several bacterial strains that are considered normally pathogens of the intestinal tract as: Escherichia coli, Enterococcus faecium, Salmonella enterica, Shigella dysenteriae, Listeria monocytogenes, Yersinia enterocolitica, Clostridium perfringens, Pseudomonas aeruginosa and Staphylococcus aureus.

Toxigenic isolate of Fusarium sporotrichioides was tested for the T-2 toxin production on PDA plates during 10 days under various glucose concentrations. T-2 toxin was determined by LC-MS and confirmed with LC-MS/MS. This analytical method has been applied, for the first time, to an extensive study of T-2 accumulation. Results showed that the production of this mycotoxin is directly correlated to the concentration of glucose present in the medium. Concentrations of T-2 toxin produced by the strain of F. sporotrichioides ranged from 0 to 1.45 mg/kg. The better T-2 production was evidenced in the fermentation operated with 20% of glucose.

Aims: Strains of Trichoderma spp. produce numerous bioactive secondary metabolites. The in vitro production and antibiotic activities of the major compounds synthesized by Trichoderma harzianum strains T22 and T39 against Leptosphaeria maculans, Phytophthora cinnamomi and Botrytis cinerea were evaluated. Moreover, the eliciting effect of viable or nonviable biomasses of Rhizoctonia solani, Pythium ultimum or B. cinerea on the in vitro production of these metabolites was also investigated. Methods and Results: T22azaphilone, 1-hydroxy-3-methyl-anthraquinone, 1,8-dihydroxy-3-methyl-anthraquinone, T39butenolide, harzianolide, harzianopyridone were purified, characterized and used as standards. In antifungal assays, T22azaphilone and harzianopyridone inhibited the growth of the pathogens tested even at low doses (1–10 μg per plug), while high concentrations of T39butenolide and harzianolide were needed (>100 μg per plug) for inhibition. The in vitro accumulation of these metabolites was quantified by LC/MS. T22azaphilone production was not enhanced by the presence of the tested pathogens, despite its antibiotic activity. On the other hand, the anthraquinones, which showed no pathogen inhibition, were stimulated by the presence of P. ultimum. The production of T39butenolide was significantly enhanced by co-cultivation with R. solani or B. cinerea. Similarly, viable and nonviable biomasses of R. solani or B. cinerea increased the accumulation of harzianopyridone. Finally, harzianolide was not detected in any of the interactions examined. Conclusions: The secondary metabolites analysed in this study showed different levels of antibiotic activity. Their production in vitro varied in relation to: (i) the specific compound; (ii) the phytopathogen used for the elicitation; (iii) the viability of the elicitor; and (iv) the balance between elicited biosynthesis and biotransformation rates. Significance and Impact of the Study: The use of cultures of phytopathogens to enhance yields of Trichoderma metabolites could improve the production and application of novel biopesticides and biofertilizers based on the active compounds instead of the living microbe. This could have a significant beneficial impact on the management of diseases in crop plants.