Agricultural and Food Science, Vol. 13 (2004): 54–67 54 © Agricultural and Food Science Manuscript received July 2003 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Contents of trichothecenes in oats during official variety, organic cultivation and nitrogen fertilization trials in Finland Veli Hietaniemi MTT Agrifood Research Finland, Chemistry laboratory, FIN-31600 Jokioinen, Finland, e-mail: veli.hietaniemi@mtt.fi Markku Kontturi MTT Agrifood Research Finland, Plant Production Research, FIN-31600 Jokioinen, Finland Sari Rämö, Merja Eurola MTT Agrifood Research Finland, Chemistry Laboratory, FIN-31600 Jokioinen, Finland Arjo Kangas, Markku Niskanen MTT Agrifood Research Finland, South Ostrobothnia Research Station, FIN-31600 Jokioinen, Finland Marketta Saastamoinen Satafood Development Association, Risto Rytin katu 70 C, FIN-32700 Huittinen, Finland Natural toxins, such as mycotoxins, have emerged as a significant factor affecting the safety image of cereal grains as a raw material for the food and feed industry. The aim of the present study was to investigate the contents of trichothecenes in representative samples of oats during official variety, nitrogen fertilization and organic farming trials in Finland, 1997–1999. Further objectives were to promote industry and commerce by selection of high-quality oat varieties for various applications. The official variety trials conducted at 8–10 locations were managed following standard protocol. There were 2 types of agronomy trial, the first included comparison of oat cultivars grown in conven- tional and organic farming systems at 6 locations, and the second used 5 nitrogen rates (0, 40, 80, 120 and 160 kg N ha-1) at 2 locations. Regardless of wet cold summer occurring in Finland during 1998, the concentrations of Fusarium toxins were lowest during this 3 year monitoring period. More myco- toxins were produced during the warm, dry summers of 1997 and 1999 than in 1998. In all, 55% of the oat samples in the official variety trials contained deoxynivalenol (DON) within the range of 50– 896 µg kg-1. The differences in DON concentrations between organic and conventional cultivation were small. The results showed also that the use of various nitrogen fertilization levels only slightly affected the trichothecene concentrations. The contents of trichothecenes in Finnish grains appeared to be similar to or lower than those reported earlier in the Northern Hemisphere. Key words: oats, grains, mycotoxins, trichothecenes, Fusarium toxins, official variety trials, nitro- gen, organic farming, cultivars mailto:veli.hietaniemi@mtt.fi 55 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Introduction The central goal of grain cultivation is the pro- duction of high-quality food or feed-related raw materials for the processing industry. Natural toxins, such as mycotoxins, have emerged as a significant factor affecting the safety image of cereal grains as a raw material for the food and feed industry. Many previous studies in Finland (Karppanen et al. 1985, Hietaniemi and Kumpu- lainen 1991, 1993, Rizzo 1993, Eskola et al. 2001, Rizzo et al. 2001) and other European countries (Tanaka et al. 1988, Langseth et al. 1989, 1999, Müller and Schwadorf 1993, Pet- tersson et al. 1995, Rizzo et al. 2001, Widestrand 2001, Döll et al. 2002, Schollenberger et al. 2002) as well as globally (Tanaka et al. 1988, Shephard et al. 1996, Groves et al. 1999, Janard- hana et al. 1999, Salay and Mercadante 2002) have shown that there is reason to focus on Fusarium toxins and their appearance. A planned European Union directive will specify the max- imum limits for trichothecenes such as 4-deox- ynivalenol (DON), T-2 toxin and HT-2 toxin and for zearalenone (ZEN) and fumonisins. DON is the most frequently found contaminant of oats, barley, wheat and corn throughout the world (Scott 1989, WHO 1993, 2001). In addition to DON, T-2 toxin, HT-2 toxin and ZEN frequent- ly occur in cereal crops cultivated in northern temperate regions (Hietaniemi and Kumpulainen 1991, Pettersson et al. 1995, Langseth et al. 1999, Rizzo et al. 2001, Thuvander et al. 2001, Eskola 2002, Lígia Martins and Martins 2002). On the other hand, fumonisins cause more extensive problems in the Southern Hemisphere than in the Northern (Shephard et al. 1996). The most fre- q u e n t l y i s o l a t e d F u s a r i u m s p e c i e s a r e F. graminearum, F. culmorum, F. moniliforme, F. poae, F. equiseti and F. proliferatum (Ylimäki et al. 1979, Marasas et al. 1984, Hietaniemi and Kumpulainen 1991, Eriksen and Alexander 1998, Eskola et al. 2001, Creppy 2002). Numerous studies on mould toxicoses have shown that type A trichothecenes such as T-2 toxin, HT-2 toxin, diacetoxyscirpenol (DAS) and neosolaniol are more acutely toxic, whereas type B trichothecenes such as DON and nivalenol (NIV) are less toxic and are therefore implicat- ed in more chronic toxicoses (Atroshi et al. 2002, Creppy 2002, Gutleb et al. 2002, Vilà et al. 2002, de Vries et al. 2003, Sudakin 2003). For exam- ple, the most frequently found of these substanc- es (DON) in cereal crops is known to affect symptoms of intoxication in humans such as abdominal pain or a feeling of fullness in the abdomen, dizziness, headache, throat irritation, nausea, vomiting, diarrhoea, and blood in the stool (Eriksen and Alexander 1998). In the case of intoxication in animals, symptoms such as loss of appetite, reduced weight gain, vomiting, heavy diarrhoea and swelling have been reported (Jof- fe 1978, Ueno 1987). In addition, DON is a very stable compound occurring during both storage and processing of food and does not degrade at high temperatures (Scott 1991). Increased attention has been focused on my- cotoxins in Finland, especially the late 1980s, Hietaniemi and Kumpulainen (1991) showed that almost all grain and feed samples studied in Fin- land during 1987–1988 contained from 7 to 300 µg kg-1 of DON and smaller amounts (13– 120 µg kg-1) of 3-acetyldeoxynivalenol. The most toxic trichothecenes, T-2- and HT-2 toxin, as well as NIV and ZEN, were found at low concentra- tions in some samples. Based on the results of Rizzo (1993) trichothecene concentrations in Finnish grains (n = 292) have ranged 1–6300 µg kg-1 for DON, 1–1000 µg kg-1 for NIV and 5– 238 µg kg-1 for T-2. In the study of Eskola et al. (2002) DON was detected in 54 of 68 Finnish cereal samples in the concentration range 5–111 µg kg-1. NIV and HT-2 toxin were detected in 3 and 2 samples, respectively, in the concentration range 10–20 µg kg-1. In addition, Eskola et al. reported the contents of Fusarium toxins in 47 cereal grain samples collected in different parts of the country. The mean concentrations of DON, 3-AcDON, NIV, HT-2 and ZEN in contaminat- ed samples were 60, 30, 42, 43 and 5 µg kg-1, respectively. T-2 was detected only once (23 µg kg-1), while fusarenon X (FX) and diacetoxy- scirpenol (DAS) were not detected. 56 A G R I C U L T U R A L A N D F O O D S C I E N C E Hietaniemi, V. et al. Contents of trichothecenes in oats There are differences in susceptibility to Fusarium contamination amongst the various types of grain. Oats are regarded as being more susceptible to Fusarium fungi in Nordic coun- tries, although studies done in Norway (Lang- seth et al. 2001) suggest that contamination in barley and wheat occurs just as frequently. The species prevalent here and in the other Nordic countries is F. avenaceum, which does not pro- duce DON and becomes especially abundant during rainy, cool summers, such as in 1998. According to Marasas et al. (1984) and Eriksen and Alexander (1998) high temperatures during the growing season favour significant toxin for- mulators; in particular, F. culmorum, F. gramine- arum and F. poae proliferate in warm tempera- tures, which was quite clearly found in Finland in the year 2001 and 2002. A belated harvest in- creases the growth of Fusarium fungi and toxin- related risk along with the rise in moisture. Fusarium fungi form toxins during the growing season, in warm-air drying, the fungi are pre- served in grain, but trichothecene compounds no longer form under dry storage. The aim of the present study during 1998– 2000 was to investigate the contents of tri- chothecenes in representative samples of oats during official variety, nitrogen fertilization and organic farming trials in Finland, 1997–1999. Further objectives were to enhance producers procedures for growing and harvesting oats and promote industry and commerce by selection of high-quality oat varieties for various applica- tions. Material and methods Samples The oat samples were collected after harvest during official and agronomy trials conducted by MTT Agrifood Research Finland in 1997–1999. The official variety trials conducted at 8–10 lo- cations were managed following standard pro- tocol (Fig. 1). There were 2 types of agronomy trial, the first included comparison of oat culti- vars grown in conventional and organic farming systems at 6 locations, and the second used 5 nitrogen rates (0, 40, 80, 120 and 160 kg N ha-1) with 4 oat cultivars at 2 locations (Fig. 1). More detailed information on the trials was published previously (Järvi et al. 2000, Eurola et al. 2003). After harvest, the grains were immediately dried with warm air in a flat bed grain drier to a mois- ture content of below 14%. The oat grains were sorted with a 2.0-mm sieve and hulled with a laboratory hulling machine BT 459 using air pressure. Oat groats were milled with a falling number hammer mill using a 1.0-mm sieve. The total number of oat samples analysed were 147, 147 and 99 in 1997, 1998 and 1999, respective- ly. The varieties studied were Leila, Kolbu, Salo, Belinda, Veli, Roope, Aarre, Katri, Puhti and Yty. Kolbu and Roope have yellow husks, and the other varieties have white husks. Leila and Kol- bu are cultivars developed in Norway, Salo and Fig. 1. Location of the trial sites in Finland. 57 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Belinda in Sweden, and Veli, Roope, Aarre, Katri, Puhti and Yty in Finland. The oat varie- ties selected for the project included the most popular cultivars as well as new varieties on the National list of cultivars in Finland. For example, the variety Veli was the most widely cultivated variety of oat in Finland from 1997 to 1999. Veli gives high quantities, is early and possesses good quality. Cultivation-related reliability is favourable and the grain is white- hulled, mid-sized and rather thin-hulled. The protein content and the weight of the grain are high. Veli is a thriving variety, reliable under cultivation, that also produces good harvests during poor years. Veli is suitable as an oat for feed, food products and export. The year of Na- tional variety catalogue entry for this type, which was developed in Finland by Boreal Plant Breed- ing Ltd, was 1982. On the other hand, Salo is a variety devel- oped in Sweden by the Svalöf Weibull Company and emerged on the market in 1989. It has good lodging resistance and was the second most widely cultivated variety of oat in Finland from 1997 to 1999. Salo is a late variety with large grain size, low hull content and pure resistance against dryness in addition to being susceptible to oat leaf blotch. Due to its short, durable stem, Salo withstands 10–20 kg ha-1 of stronger nitro- gen fertilization than normal. The main use of Salo has been as a food product. Analytical methods Sample clean-up, identification and quantification To determine the presence of mycotoxins, the ground-up samples (25 g) were extracted through a blend of acetonitrile-water (84:16) and then suction-filtered. Part of the filtrate (7.5 ml) was extracted for activated carbon cleaning (My- coSep#227, Romer Labs), after which the sam- ple was transferred to a silylated test tube and evaporated to dryness under a stream of nitro- gen. 100 µl N-trimethylsilylimidazole (TMSI) reagent was added to the residue evaporated to dryness. DON, DAS, 3-acetyldeoxynivalenol (3- AcDON), FX, NIV, T-2 and HT-2 toxins and 19- nortestosterone (the internal standard) were iden- tified and quantified as trimethylsilylether de- rivatives, employing GC-MS (Rizzo et al. 1986, Hietaniemi and Kumpulainen 1991, Saastamoi- nen and Saloniemi 1997). Quality assurance The reliability of the GC-MS method in tri- chothecene analysis was examined in repeata- bility, reproducibility, recovery and intercalibra- tion studies by specifying the linearity of the method, the minimum detectable concentrations (quantification limit), utilizing certified refer- ence materials and participating in FAPAS round tests. The recoveries for the compounds varied between 70–100% and the repeatability between 5% and 25%, while the minimum detectable con- c e n t r a t i o n s w e r e 5 0 µg k g - 1 w i t h a l l t r i - chothecenes. The various linearity areas with respect to trichothecenes were as follows: DON (25–8000 µg kg-1), DAS (25–500 µg kg-1), 3- AcDON (25–800 µg kg-1), FX (25–250 µg kg-1), NIV (25–400 µg kg-1), T-2 (25–600 µg kg-1) and HT-2 (25–1300 µg kg-1). BCR wheat flour, CRM 379 was employed as the reference material (DON concentration 670 µg kg-1). The Chemis- try Laboratory of Agrifood Research Finland follows a quality control system in accordance with ISO 17025. In the case of samples whose concentration level was below the limit of quan- tification, the value reported in the mean value, median and standard deviation calculations was estimated as 50% of the quantification limit. For statistical analyses Data Desk 6.1.1 (Data exploration and visualization) was used (Velle- man and Hoaglin 1981). Results and discussion Official variety trials The results of the official variety trials showed that the mycotoxin DON was found most fre- 58 A G R I C U L T U R A L A N D F O O D S C I E N C E Hietaniemi, V. et al. Contents of trichothecenes in oats quently in Finnish oats during 1997–1999. In 1997, the procent of positive toxin findings out of all oat samples studied (51) were as follows: DON 69%, DAS not detected, 3-AcDON 6%, FX not detected, NIV 14%, T-2 toxin 4% and HT-2 toxin 4%. The mean DON concentration of all samples was 222 µg kg-1 (median 79 µg kg-1, standard deviation 206 µg kg-1). A few individu- al, rather high contents of NIV were found within a range of < 50–575 µg kg-1. The mean T-2- and HT-2 toxin concentrations were below the quan- tification limit of 50 µg kg-1. The highest mean DON concentrations were determined in the ar- eas of Hyrylä, Pälkäne, Mikkeli and Laukaa (Fig. 1). The mean contents of DON, amount of precipitation, effective temperature sum values, and pH and the type of soil as obtained for May – August are presented in Table 1. None of the above-mentioned background factors did not explain directly the higher toxin concentrations in these areas. The amounts of precipitation and effective temperature sums were similar to the average values observed in other areas, and not even the soil types could indicate a direct con- nection with elevated toxin concentrations (Ta- ble 1). The varieties, Leila, Salo, Veli, Belinda, Kolbu and Roope, were employed in the official trials. Figure 2 shows DON contents of six oat Table 1. Precipitation, effective temperature sum, soil pH, soil type and mean deoxynivalenol (DON) content of oats in official variety trials during 1997–1999 at different trial locations in Finland. Location Year Precipitation Effective Soil pH Soil type Mean DON May-Augmm temperature sum content µgkg-1 Jokioinen 1997 302 1217 6.3 sandy clay 64 1998 318 1011 5.8 clay < 50 1999 146 1184 5.7 sandy clay 243 Mietoinen 1 1997 232 1285 5.3 clay 87 1998 245 1036 5.9 sandy clay 97 1999 92 1236 5.8 sandy clay 262 Hyrylä 1997 not available not available 6.3 sandy clay loam 630 1998 not available not available 6.0 coarse silt 114 Pälkäne 1997 253 1250 5.7 fine silt 334 1998 339 1043 6.0 fine silt < 50 1999 141 1231 5.9 fine silt 184 Mikkeli/Juva 1997 115 1150 6.9 fine sand 272 1998 337 976 6.0 fine sand < 50 1999 243 1133 6.0 fine sand < 50 Maaninka 1997 177 1153 5.8 coarse silt 233 1998 340 938 6.1 coarse silt 165 1999 183 1134 5.5 coarse silt 286 Laukaa 1997 182 1146 6.0 coarse silt 435 1998 345 916 6.0 coarse silt 419 1999 175 1112 6.0 coarse silt 88 Ylistaro 1 1997 155 1143 6.0 sandy clay loam < 50 1998 372 937 6.2 silty clay < 50 1999 120 1062 6.1 silty clay < 50 Ylistaro 2 1997 155 1143 5.3 mould < 50 1998 372 937 5.7 mould < 50 1999 120 1062 5.5 mould 82 Ruukki 1 1999 182 952 5.4 coarse silt 102 Ruukki 2 1999 182 952 5.4 mould < 50 Vihti 1998 389 997 5.9 coarse silt 70 1999 132 1173 6.2 clay 260 59 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Fig. 2. Deoxynivalenol (DON) contents of six oats cultivars across all sites of official variety trials during 1997–1999 in Finland. 60 A G R I C U L T U R A L A N D F O O D S C I E N C E Hietaniemi, V. et al. Contents of trichothecenes in oats cultivars across all sites of official variety trials during 1997–1999. The highest individual DON concentrations were found in the varieties Veli and Salo. According to the statistical analyses no distinct differences were found in DON con- tents of various varieties. It is most likely that the climate had the greatest impact by fixing the time of heavy rainfalls in the right period of the growing season, for example during heading and harvesting time. During these 3 research years, the lowest mycotoxin concentrations were determined from the oat harvest of 1998 (Table 2). The mean DON content from the 51 samples studied was 99 µg kg-1 (median < 50 µg kg-1, standard deviation 171 µg kg-1). Other toxins were found in only a few samples as small concentrations, e.g., no T- 2 toxin content exceeding the quantification limit was found and only one HT-2 toxin concentra- tion in excess of the quantification limit was de- termined. Summer 1998 was cool and rainy throughout the Finland. Despite this, conditions for the formation of toxins were not favourable. Nevertheless, fusarium head blight visibly ap- peared in Finnish grain, especially in rye. In the investigations of Eskola et al. (2000 and 2001) a lot of mouldiness of the Finnish grain in 1998 was found. Despite of the heavy contamination of the moulds, contents of trichothecenes were low. The findings in 1998 also indicated that Veli and Salo had the highest individual DON con- centrations. The highest DON concentrations were detected in Laukaa, central Finland, where rainfall was also high and the effective tempera- ture sum was low. However, similar amounts of rain were also observed in other areas, and there was no association with elevated toxin concen- trations within the same area (Table 1). As a research year 1999 was quite similar to 1997 (Table 2). In several areas, 1999 was clear- ly less subject to rain, but the effective tempera- ture sums were similar to those in 1997. The average DON concentrations for the oat samples were, 154 µg kg-1, and the number of positive findings out of the 59 samples studied was 37. The largest DON concentrations were detected in the variety Veli. As in 1998, no positive T-2 toxin results were measured, but 4 positive HT- 2 toxin contents were found, the highest value being 240 µg kg-1. NIV was found from 14 sam- ples out of a total number of 59, with the con- centrations ranging < 50–423 µg kg-1. From the regional stand point, the most contaminated oat samples were found in southern, southwestern and eastern Finland (Fig. 1). The most precipi- tation occurred in eastern Finland within a range of 183–243 mm, and the least was measured in southwestern Finland at 92 mm. The year 1999 also indicated that a direct connection between the amounts of rain, effective temperature sums and contents of mycotoxins does not exist. It still appears more probable that the ‘right’ period of rainfall during the growing season ex- erts greater significance. For example, if heavy rainfalls occurred during heading and harvest- ing time, the risk of growth of Fusarium fungi and production of toxins would exist. Oldenburg et al. (2000) studied the factors which influence most the infection of grain, especially wheat. The authors found the climate to have the greatest impact, followed by infection pressure/tillage, corn as preceding crop, plant protection, culti- vars and plant nutrition. Langseth et al. (2001) reported from Norway that the DON level was especially high in 1988 and 1992, years with spring drought and much precipitation in July in the period during and after anthesis. According to McMullen et al. (1997) during the last two decades fusarium head blight disease occurs es- pecially during rainy flowering seasons of the crop and besides of DON, an infection of the head by Fusarium can lead to severe yield loss and reduced kernel quality. On the other hand, rains occurring at harvest time may increase the risk of appearance of mycotoxins considerably. How- ever, warm-air-drying as primarily employed in Finland is evidently a security-enhancing factor with respect to grain (Jayas and White 2003). Nitrogen fertilization trial Nitrogen fertilization trials at five nitrogen lev- els 0, 40, 80, 120 and 160 kg N ha-1 were imple- 61 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Table 2. Mean contents of trichothecenes in oats during official variety trials in Finland, 1997-1999. Number of samples Range µgkg-1 Mean µgkg-1 Median µgkg-1 StdDev µgkg-1 Positive [%] 1997 DON 51 < 50–706 222 79 206 69 DAS 51 Not detected 3-AcDON 51 < 50–219 < 50 < 50 41 6 F-X 51 Not detected NIV 51 < 50–575 74 < 50 129 14 T-2 51 < 50–349 < 50 < 50 62 4 HT-2 51 < 50–507 < 50 < 50 85 4 1998 DON 52 < 50–896 99 < 50 171 33 DAS 52 Not detected 3-AcDON 52 < 50–310 < 50 < 50 50 10 F-X 52 Not detected NIV 52 < 50–530 53 < 50 95 10 T-2 52 Not detected HT-2 52 < 50–116 2 1999 DON 59 < 50–660 154 108 158 63 DAS 59 Not detected 3-AcDON 59 Not detected F-X 59 Not detected NIV 59 < 50–423 < 50 < 50 63 24 T-2 59 Not detected HT-2 59 < 50–240 < 50 < 50 < 50 7 mented. Aarre, Katri, Kolbu and Salo were the varieties used in the trials, which were carried out in 2 locations: in Jokioinen, province of Häme and Ylistaro, South Ostrobothnia (Fig. 1). The results showed that the use of various nitro- gen fertilization levels during 1997–1999 only slightly affected the mycotoxin concentrations (Fig. 3). The highest mean DON concentrations were observed in the data from 1999 within a range of 101–730 µg kg-1 (Fig. 3). The DON con- centrations in Häme (Jokioinen) were, to some extent, higher than those in South Ostrobothnia (Ylistaro), although the amounts of rainfall were similar. No remarkable differences between the various varieties were found; the highest indi- vidual concentrations were determined from the variety Aarre (Fig. 3). Aarre is an early, high- yielding and high-quality oat variety and in ad- dition has good lodging resistance. It is earlier than Veli by one day. Aarre is used for feed and food products and export and it is developed by Boreal Plant Breeding Ltd of Finland. It was in- troduced in the National variety catalogue in 1995. Organic farming The trials for conventional as well as organic cultivation were implemented during 1997–1998 at 6 locations: Jokioinen, Mietoinen, Laukaa, Partala, Ylistaro and Ruukki. Only at the Partala Research Station in eastern Finland was conven- tional cultivation not used as comparative data. More precise comparison of these cultivation methods could be carried out only with 2 varie- ties, Veli and Puhti, although 5 other varieties Aarre, Katri, Kolbu, Leila and Roope were also included in the organic cultivation trials. In con- trast to the results obtained in the official varie- ty trials and nitrogen fertilization trials, DON was also the most commonly appearing myco- 62 A G R I C U L T U R A L A N D F O O D S C I E N C E Hietaniemi, V. et al. Contents of trichothecenes in oats Fig. 3. Mean deoxynivalenol (DON) contents at five nitrogen levels in nitrogen fertilization trials in 1999 in Finland at two sites (Jokioinen and Ylistaro). 63 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. Fig. 4. Mean deoxynivalenol (DON) contents in oats during organic and conventional cultivation trials in Finland in 1997 and 1998. 64 A G R I C U L T U R A L A N D F O O D S C I E N C E Hietaniemi, V. et al. Contents of trichothecenes in oats toxin in samples from the conventional and or- ganic cultivation trials (Fig. 4). Corresponding- ly, the results corroborated previous indications that the toxin concentrations during the rainy, cold year of 1998 were clearly lower than in 1997. There were no large differences in DON concentrations in the cultivation methods stud- ied. The mean levels appeared to be similar in organic and conventional cultivation (Fig. 4). According to Leblanc et al. (2002) a particu- lar group at risk for exposure to DON may be the consumers of organic products as they may regularly eat foods containing higher levels of DON than the general population. On the basis of the work of Schollenberger (2002) the DON content of flour samples originating from con- ventionally produced wheat was significantly higher than that of samples from organic pro- duction. Similar results were obtained by Döll et al. (2000) and Usleber et al. (2000), who found lower DON contents in wheat, rye and flour sam- ples of organic origin than in those from con- ventional production. In agreement with this study, Marx et al. (1995) reported from south- ern Germany that the incidence of DON in con- ventionally and organically produced wheat was 88% and 76%, whereas the mean contents were 420 and 486 µg kg-1, respectively. In addition, according to Döll et al. (2000) the organic farm- ing system with the practice of a well-balanced crop rotation, tillage and fertilization showed benefits concerning the contamination with Fusarium toxins, especially DON. Conclusions The results of the official variety and nitrogen fertilization trials and comparison of convention- al and organic cultivation, suggest that future research and follow-up on trichotecenes must be emphasized and continued. The results of the trials showed that the mycotoxin DON was found most frequently in Finnish oats during 1997– 1999. According to the results on an average 55% of the oat samples respective to the official va- riety trials in 1997–1999 contained DON within the range of 50–896 µg kg-1. Correspondingly, the range and frequencies of other toxin find- ings were as follows: 3-AcDON 50–310 µg kg-1 (5%), NIV 50–575 µg kg-1 (16%), T-2 toxin 50– 349 µg kg-1 (1%) and HT-2 toxin 50–507 µg kg- 1 (4%). A directive currently under preparation in the European Union will specify the tolerance limits for DON (1500 µg kg-1). In comparison to previous studies, the contents of trichothecenes in grains appeared similar or lower to those re- ported earlier in the Northern or Southern Hem- isphere (Karppanen et al. 1985, Tanaka et al. 1988, Hietaniemi and Kumpulainen 1991, Müller and Schwadorf 1993, Rizzo 1993, Pettersson et al. 1995, Groves et al. 1999, Janardhana et al. 1999, Döll et al. 2000, Langseth and Rundber- get 2001, Rizzo et al. 2001, Eskola 2002, Schol- lenberger et al. 2002, Salay and Mercadante 2002). The results also showed that no distinct differences were found in DON contents of var- ious varieties. The differences in DON concen- trations between organic and conventional cul- tivation were small. In addition, the results showed that the use of various nitrogen fertili- zation levels only slightly affected the tricho- thecene concentrations. Nevertheless, the importance of background factors with respect to samples is often forgot- ten in monitoring the quality of grains, not to mention a deeper familiarity with their impact and, through the same, better control over grain quality. Evidently, the incidence of rain during heading time represents a risk factor (McMul- len et al. 1997, Döll et al. 2000, Oldenburg et al. 2000, Langseth et al. 2001), although it has not been indicated here. The present results also showed that more precise research into the ef- fects of cultivation methods in relation to fungi and toxins is necessary (Norred 2000). Good fa- miliarity with, the effects of the preceding crop, rotation, seed purity, various soil preparation methods, direct sowing and pesticides on the formation of mycotoxins is a minimal require- ment. A question of its own right also concerns breeding for resistance against Fusarium fungi 65 A G R I C U L T U R A L A N D F O O D S C I E N C E Vol. 13 (2004): 54–67. and their associated toxins (de Vries 2000, Hol- lins et al. 2003). Knowledge of these factors is a prerequisite for good cultivation-related direc- tives that industry and farmers should follow to ensure high-quality production of oats both in Finland and internationally. Acknowledgements. The authors wish to thank Ms. Kirsi Puisto, Ms. Leena Holkeri, Mr. Seppo Nummela and Mr. Tauno Koivisto for their skilful technical assistance and enthusiasm for this study. This study was financially sup- ported by the Ministry of Agriculture and Forestry and the Finnish food and feed industry: Raisio Group, Suomen Vil- java Ltd. and Kemira Agro Ltd. References Atroshi, F., Rizzo, A., Westermarck, T. & Ali-Vehmas, T. 2002. Antioxidant nutrients and mycotoxins. Toxicol- ogy 180: 151–167. Creppy, E.E. 2002. 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SELOSTUS Kauran trikotekeenipitoisuus virallisissa lajikekokeissa sekä typpilannoitus- ja luomulajikekokeissa Veli Hietaniemi, Markku Kontturi, Sari Rämö, Merja Eurola, Arjo Kangas, Markku Niskanen ja Marketta Saastamoinen MTT (Maa- ja elintarviketalouden tutkimuskeskus) ja Satafood Kehittämisyhdistys ry Viljojen luontaiset toksiinit, kuten hometoksiinit, ovat tärkeitä raaka-aineen laadun mittareita elintarvike- ja rehuteollisuuden kannalta. Tutkimuksessa määritettiin Fusarium-toksiinit ja trikotekeenit satokausien 1997– 1999 virallisten lajike-, typpilannoitus- ja luomula- jikekokeiden kauranäytteistä. Lisäksi tavoitteena oli lisätä tietoa eri kauralajikkeiden kemiallisesta koos- tumuksesta. Viralliset lajike-, typpilannoitus- ja luo- mulajikekokeet toteutettiin MTT:n tutkimusasemilla. Virallisten lajikekokeiden tulosten perusteella homemyrkyistä deoksinivalenoli (DON) esiintyi useimmiten suomalaisessa kaurassa vuosina 1997– 1999. Tutkituista näytteistä 55 % sisälsi DON:a < 50– 896 µg kg-1. Pienimmät trikotekeenipitoisuudet kol- men tutkimusvuoden aikana määritettiin vuoden 1998 kaurasadosta. Satokausi 1998 oli hyvin sateinen, kos- tea ja kylmä. Typpilannoituskokeissa käytettiin nel- jää eri typpitasoa 40, 80, 120 ja 160. Lisäksi vertai- luaineistona oli nollataso, jossa lisätyppeä ei käytet- ty ollenkaan. Tulosten perusteella typpilannoitusta- so ei vaikuttanut kauran mykotoksiinipitoisuuksiin vuosina 1997–1999. Suurimmat keskimääräiset DON-pitoisuudet havaittiin vuoden 1999 aineistosta vaihteluvälillä 192–510 µg kg-1. Vastaavasti kuin edel- lä esitetyt tulokset virallisista lajikekokeista ja typ- pilannoituskokeista osoittivat, deoksinivalenoli oli yleisimmin esiintyvä Fusarium-toksiini myös tavan- omaisen ja luomuviljelykokeiden näytteissä. Tutkit- tu viljelymenetelmä ei vaikuttanut DON-pitoisuuk- siin. Yhteenvetona voidaan todeta, että viljoissa esiin- tyviä trikotekeenejä on tärkeä tutkia lisää, ja järjes- telmä pitoisuuksien seurantaan on luotava. EU:ssa valmisteilla oleva direktiivi tulee määrittämään suu- rimmat sallitut pitoisuudet deoksinivalenolille, T-2- ja HT-2-toksiinille, tsearalenonille ja fumonisiineil- le. Viljanäytteiden taustatietojen tunteminen ja niiden vaikutusten analysointi nousee yhä tärkeämmäksi laa- dun seurannassa. Tämä mahdollistaa nykyistä parem- man viljan laadun hallinnan. Contents of trichothecenes in oats during official variety, organic cultivation and nitrogen fertilization trials in Finland Introduction Material and methods Results and discussion Conclusions References SELOSTUS