Barbara Birzele

Deoxynivalenol and ochratoxin A in German wheat and changes of level in relation to storage parameters

The occurrence of the mycotoxins deoxynivalenol (DON) and ochratoxin A (OTA) in the winter wheat of 1997 and 1998 grown under organic farming conditions was investigated using ELISAs (R-Biopharm®) for quantification. The influence of delayed drying of the grain after harvest on the development of DON and OTA was determined in storage trials (moisture: 17% and 20%; temperature: 20°C; duration: four and six weeks). The Tox5 PCR assay was used both to detect Fusarium species with the potential to produce trichothecenes and as a measure of their relative DNA content during the storage trials. The intensity of the PCR signals was correlated with the DON concentration. Fusarium species were identified microscopically by standard methods. All the freshly harvested grain samples were contaminated with DON and showed further increases in the DON concentration during storage. OTA contamination was found in 14.3% of the 1997 samples and in 24.1% of the 1998 samples. OTA increased during storage trials of the 1997 samples but not in the 1998 samples.

Epidemiology of Fusarium Infection and Deoxynivalenol Content in Winter Wheat in the Rhineland, Germany

Details of our long-term research programme concerning the epidemiology of Fusarium spp. and mycotoxin production are summarized. Evaluation of the occurrence of Fusarium spp., mainly on winter wheat (Triticum aestivum), was carried out by investigating Fusarium infection and mycotoxin contamination. Two to 15% of grains were infested during 1995–1998 at three climatologically differing localities of the Rhineland, Germany. Disease progress was accelerated by rainfall during the flowering season. The species most frequently isolated were Fusarium avenaceum, F. poae, F. culmorum and F. graminearum. The mean deoxynivalenol (DON) content varied from 19μgkg−1 (1995) to 310μgkg−1 (1998) and was not always correlated with disease severity. Organic farming systems showed lower rates of infection with ear blight and lower mycotoxin contamination than conventional farming systems.

Impact of growth conditions on the occurrence ofFusarium spp. and the mycotoxin content of wheat

In 1997–99 the occurrence ofFusarium spp. on winter wheat and the contamination with mycotoxins was investigated at three locations in the Rhineland, Germany. All cultivation methods investigated had an effect on the level ofFusarium infection, however, rainfall during flowering was the most important factor. The choice of cultivar and soil cultivation proved to be the most promising tools to reduce head scab severity and mycotoxin contamination.ZusammenfassungVon 1997–99 wurde das Auftreten vonFusarium spp. sowie die Belastung von Weizen mit Mykotoxinen an drei Standorten im Rheinland untersucht. Alle pflanzenbaulichen Maßnahmen konnten zu einer Reduktion der Befallshöhe führen, entscheidend für den Befall war aber die Witterung während der Blüte. Sortenwahl und Bodenbearbeitung erwiesen sich als am wirksamsten, den Ährenbefall und die Mykotoxinbelastung zu reduzieren.

Rheological and breadmaking properties of wheat samples infected withFusarium spp.

Rheological and breadmaking properties of untreated and suboptimally stored wheat samples (grain moisture: 20%, temperature: 20°C) and also of wheat which was inoculated withFusarium spp. were investigated. The deoxynivalenol (DON) content of the stored and inoculated wheat samples ranged between 820–12,000 μg/kg. Gluten proteins were isolated with different extraction solutions and the fractions obtained were analysed by means of RP-HPLC. Microextension tests and micro-baking tests were used for the determination of dough properties (maximum resistance (MR) and extensibility (EX)) and bread volume, respectively. In spite of the extremely high DON concentrations of some wheat samples contaminated withFusarium spp. they showed only a slight decrease of the amount of gluten proteins. Extension tests of dough led to a slight decrease of MR, bread volumes stayed almost the same compared with the non-contaminated grain. The contamination of wheat withAspergillus andPenicillium led to a high decrease of gluten proteins, which resulted in an extremely decreased MR of the dough and a very low bread volume.

Fusarium spp. and storage fungi in suboptimally stored wheat: Mycotoxins and influence on wheat gluten proteins

When wheat is stored under suboptimal conditions, a further mycotoxin increase of deoxynivalenol (DON), but especially of mycotoxins produced by storage fungi, e.g. ochratoxin A, is possible, lowering wheat quality and food safety. Different storage trials were conducted under suboptimal storage conditions.Fusarium survival during suboptimal storage was monitored by cultural technique and multiplex-PCR and set into relation to DON contents. Furthermore, XANES spectroscopy was applied on a selected storage trial in order to characterize sulfur speciation in low molecular weight (LMW) subunits of glutenin isolated from suboptimally stored wheat samples highly infected withFusarium and from wheat infected withAspergillus andPenicillium. Distinct changes in sulfur speciation were observed in grains infected with storage fungi, especially a significant increase of higher oxidation states (sulfoxide state, sulfonate state).

Significance of different inoculum sources for the Fusarium infection of wheat ears

From 1997 to 1999 the occurrence ofFusarium spp. on wheat grain and its contamination with the mycotoxins deoxynivalenol and invalenol were investigated under organic farming conditions in the Rhineland, Germany. For comparison, some trials were also run under integrated farming conditions. The importance of the seed contamination withFusarium spp. as well as the impact of farming system, previous crop and soil preparation on the inoculum sources ofFusarium spp. in the soil were investigated. The data on the inoculum sources was compared to the Fusarium infection of grains and their content of DON and NIV. The crop residues in the soil were the most important inoculum source for the Fusarium species infecting wheat ears and grains. The amount of potential inoculum in the soil largely depended on the previous crop and the system of soil preparation.

Development of deoxynivalenol contents in relation to the PCR detection of potentially trichothecene producingFusarium spp. during storage of wheat

Development of deoxynivalenol (DON) in wheat with a low contamination withFusarium spp. was investigated under suboptimal storage conditions (17% and 20% grain moisture, 20°C). The influence of storage on the relative DNA content of potential DON producers was also determined. The DON contents were quantified using an ELISA. The Tox5 PCR was used for the detection of potential trichothecene producers and for the estimation of their relative DNA content. ThegaoA gene was subsequently amplified by PCR to detect specificallyFusarium graminearum. The concentration ofF. graminearum DNA was semiquantitatively determined using a Light Cycler™. The DON concentrations increased during storage trials but the intensity of PCR signals decreased.ZusammenfassungDie Bildung von Deoxynivalenol (DON) in Weizen wurde, ausgehend von einem geringen Besatz mitFusarium spp., unter suboptimalen Lagerbedingungen (17% und 20% Kornfeuchte, 20°C) untersucht. Der Einfluss der Lagerung auf die relative DNA-Masse potentieller DON-Bildner wurde ebenso ermittelt. Die DON-Gehalte wurden mittels ELISA quantifiziert. Die Tox5-PCR wurde zur Detektion potentieller Trichothecen-Bildner und zur Abschätzung ihrer relativen DNA-Masse eingesetzt. DasgaoA-Gen wurde anschließend mit der PCR amplifiziert, um spezifischFusarium graminearum nachzuweisen. Die Konzentration derF. graminearum-DNA wurde im Light Cycler™ semiquantitativ bestimmt. Die DON-Konzentrationen nahmen im Verlauf der Lagerungsversuche zu, während die Intensität der PCR-Signale abnahm.