Natural occurrence of Fusarium toxins in soy food marketed in Germany

A total of 45 samples of soy food including whole beans, roasted soy nuts, flour and flakes, textured soy protein, tofu, proteinisolate including infant formulas and fermented products (soy sauce) were randomly collected in food and health food stores and analysed for Fusarium toxins. A spectrum of 13 trichothecenes of the A-type as well as of the B-type were determined by gas chromatography/mass spectrometry, zearalenone (ZEA), alpha- and beta-zearalenol (alpha- and beta-ZOL) by high performance liquid chromatography (HPLC) with fluorescence and UV-detection. Detection limits ranged between 1 and 19 microg/kg. At least one of the toxins investigated was detected in 11 out of a total of 45 samples of soy food belonging to different commodities. Scirpentriol (SCIRP), 15-monoacetoxyscirpenol, 4,15-diacetoxyscirpenol, T-2 tetraol, HT-2 toxin, deoxynivalenol (DON), 15- and 3-acetyldeoxynivalenol, ZEA, alpha- and beta-ZOL were detected in at least one sample, T-2 triol, T-2, NEO, NIV and FUS-X were not detected in any sample. Five out of 11 samples were positive for one toxin, one sample for two, three, six or seven toxins, two samples for 5 toxins, demonstrating the possibility of a contamination of soy food with a spectrum of Fusarium toxins. SCIRP, DON and ZEA were found up to 108, 260 and 214 microg/kg, the other toxins did not exceed 61 microg/kg. A first insight into the contamination of soy food with a broad spectrum of Fusarium toxins is provided.     

Fusarium toxins in wheat flour collected in an area in southwest Germany

A total of 60 samples of wheat flour were collected during the first 6 months of 1999 from mills and food stores in an area in southwest Germany. Samples included whole-grain and two types of white flour with these three groups characterized by a high, medium and low ash content. The contents of deoxynivalenol (DON), nivalenol (NIV), 3- and 15-acetyldeoxynivalenol, HT-2 toxin (HT-2), T-2 toxin (T-2) and fusarenon-X (FUS-X) were determined by gas chromatography/mass spectrometry, and those of zearalenone (ZEA), alpha- and beta-zearalenol (alpha- and beta-ZOL) by high performance liquid chromatography with fluorescence detection. FUS-X, alpha- and beta-ZOL were not detected in any sample. Based on incidence and level, DON was the predominant toxin followed by NIV and ZEA for all three flour types. The overall degree of toxin contamination was lower with decreasing ash content. This suggests a localization of the toxins analyzed primarily in the outer parts of the original wheat kernels. The median DON content was significantly (P<0.05) higher for wheat flour originating from wheat of conventional than of organic production.     

Natural occurrence of Fusarium toxins in oats harvested during five years in an area of southwest Germany.

A total of 56, 56, 54, 51, and 55 oats samples used for feed production were collected randomly after the 1987, 1989, 1990, 1991 and 1992 crops, respectively, from farms located in an area of southwest Germany. Deoxynivalenol (DON), 3- and 15-acetyl-deoxynivalenol (3-, 15-ADON), nivalenol (NIV), fusarenon-X (FUS-X), T-2 toxin (T-2), HT-2 toxin (HT-2) and diacetoxyscirpenol (DAS) were determined by gas chromatography with mass selective detection (GC-MS), zearalenone (ZEA), alpha and beta-zearalenol (alpha-, beta-ZOL) by GC-MS or by HPLC. DON was the major toxin with incidences at 49-85% and mean levels in positive samples of 52-302 micrograms/kg. Incidences of ZEA, 3-ADON, NIV, HT-2, and T-2 were at 20-37, 0-30, 18-67, 0-29, and 27-61%, respectively, with mean levels in positive samples at 8-25, 5-63, 11-192, 205-296, and 20-244 micrograms/kg, respectively. alpha- and beta-ZOL and DAS were not detected in any sample. 15-ADON and FUS-X were assayed in samples from 1987, 1991 and 1992. 15-ADON was detected in 9, 4 and 0% of samples, with an average of 9 and 18 micrograms/kg, respectively; FUS-X was not detected. The incidence and levels of toxins varied from year to year. The correlation between the occurrence of toxins and precipitation is discussed.     

Protein inhibitors of hydrolases in plant foodstuffs

Abstract

In this review protein inhibitors of hydrolases occurring in plant tissues used as foods are treated, that is, proteins inhibiting digestive enzymes such as proteases (trypsin and chymotrypsin, which are discussed in more detail, elastase, pepsin, carboxypeptidases), amylases, lipases, and some other hydrolases. Topics covered are the chemistry of these inhibitors including their isolation, structure, and properties; their physiological function; their occurrence in plants and foodstuffs, with special emphasis on their inactivation during processing; their toxicology; and the uptake by human beings. In addition, analytical aspects are discussed in greater detail, and procedures for inhibitor activity determination are included.     

Contamination of some foodstuffs marketed in Italy by fatty acid esters of monochloropropanediols and glycidol

ABSTRACT

Fatty acid esters of 3-monochloropropanediol (3-MCPD), 2-MCPD and glycidol (Gly) are food-processing contaminants that cause concerns about possible adverse health effects. The present study evaluates the contamination levels of the three ester classes in 130 samples of foodstuffs marketed in Italy covering 10 food categories, namely margarines, oils, roasted coffee, breakfast cereals, salted crackers, cookies, infant biscuits, rusks, breads and potato crisps. The analytical method employed is a so-called indirect method that entails MCPD/glycidol cleavage from their esterified forms, cleanup, derivatisation and GC-MS analysis. The MCPDs and glycidol concentrations (from esters) were found to be equal or a little higher than the levels reported in previous studies conducted in other European countries and described in the literature. 3-MCPD was the predominant compound in all foodstuffs analysed with the exception for rusks where Gly levels were slightly higher. Considering the sum of MCPD and Gly esters, the most contaminated foodstuffs were seed oils, followed by margarines and cookies, whereas roasted coffee, bread, rusks, cornflakes and infant biscuits were less contaminated with MCPDs and Gly concentrations often below LOQ or LOD values. Refined olive oil, potato chips and salted crackers showed contamination levels intermediate between the two above groups. The results of this study also confirm that the use of palm oil as an ingredient or frying medium is an important cause of increase of the levels of MCPD and Gly esters, especially in salted crackers, rusks and potato crisps. Finally, the Italian intake of 3-MCPD due to the various foods analysed has been calculated and related to TDI. The MoE for Gly was also estimated.     

Natural co-occurrence of deoxynivalenol and fumonisins B1 and B2 in Italian marketed foodstuffs

A survey was performed to obtain the frequency and levels of contamination by deoxynivalenol (DON) and fumonisins B₁ and B₂ (FB1, FB2) mycotoxins in Italian marketed foods. Of 202 samples investigated, including raw materials and processed cereal foods (bread, pasta, breakfast cereals, biscuits, baby and infant foods), 84% were contaminated with DON at levels from 0.007 to 0.930 μg g^-1 (median 0.065 μg g^-1); 26% contained FB1 ranging from 0.010 to 2.870 μg g^-1 (0.070 μg g^-1); 35% contained FB2 at 0.010-0.790 μg g^-1 (0.080 μg g^-1). The highest levels of DON and FB1 were detected in raw cereals and wholemeal flours. The highest levels of FB2 were detected in durum wheat pasta. A widespread DON contamination was found in baby and infant foods at levels varying from 0.007 to 0.166 μg g^-1.     

Naturally occurring Fusarium toxins in New Zealand maize

Twenty samples of maize collected from healthy growing crops and at harvest time and during storage were screened for four Fusarium toxins (deoxynivalenol, diacetoxyscirpenol, T-2 toxin and zearalenone) by gas chromatography-mass spectrometry and thin-layer chromatography. Seventeen samples (85%) contained one or more of these toxins. Zearalenone was present in 15 samples at levels ranging between 0.1 and 16 ppm. Deoxynivalenol, diacetoxyscirpenol and T-2 toxin were found in 11, 6 and 13 samples respectively, all at levels below 1 ppm. This work documents the first reported natural occurrence of Fusarium toxins in New Zealand maize, and it is concluded that all four of the mycotoxins studied are prevalent in apparently healthy standing crops as well as in stored maize.     

Occurrence of Fusarium T-2 and HT-2 toxins in oats from cultivar studies in Germany and degradation of the toxins during grain cleaning treatment and food processing

For the Fusarium trichothecene mycotoxins T-2 and HT-2, a combined (T-2 + HT-2) temporary tolerable daily intake (tTDI) of 0.06 µg kg^?1 body weight day^?1 was proposed at the European level in 2001 (Opinion of the Scientific Committee on Food). In the near future, maximum levels for these trichothecenes will be regulated by the European Commission as announced in EU (VO) 1881/2006. For the implementation of these maximum levels, more data on occurrence and behaviour of T-2 and HT-2 toxins in primary agricultural products as well as during cleaning treatment and food processing are needed. In the current work, we determined the T-2/HT-2 concentrations in four oat cultivars (Aragon, Dominik, Ivory, Pergamon) from ten different agricultural sites in Germany, grown in cultivar studies in 2007. The grains were de-hulled, oat meal was prepared, and bread with 20% oat meal and 80% wheat flour was baked. In the cereal-processing chain, samples were taken at various steps and subsequently analysed for their T-2/HT-2 content. We employed liquid chromatography-mass spectrometry (LC-MS) and an immunological screening method (enzyme-linked immunoabsorbant assay (ELISA)) for T-2/HT-2 determination. Detection limits were between 1 and 10 µg kg^?1 in different matrices. T-2/HT-2 concentrations determined by ELISA in oat samples from ten different agricultural sites in Germany were between 9 and 623 µg kg^?1. The median and 90th percentile were 48 and 191 µg kg^?1 T-2/HT-2, respectively. One site showed six times higher T-2/HT-2 levels than the other sites, where concentrations ranged from 322 to 623 µg kg^?1. In 80% of the samples the cultivars Pergamon and Ivory had the lowest concentration of T-2 and HT-2 toxins. Using LC-MS for T-2/HT-2 determination, cleaning of the raw material did not lead to significant reductions of T-2 and HT-2 levels, whereas de-hulling led to a reduction of over 90%. Boiling of oat meal produced from cleaned raw material to yield ‘porridge’ resulted in varying T-2/HT-2 levels in experimental replicates. No major reduction of T-2/HT-2 levels in cooked porridge was obtained. Standardized baking experiments using 20% oat meal showed that T-2 and HT-2 toxins are relatively stable during the baking process, probably due to their temperature stability.     

Survey of the furosine content in cheeses marketed in Spain

The furosine content in 53 samples of different cheese types (fresh, ripened with molds, artisanal hard-pressed, industrial hard-pressed, and processed) marketed in Spain was analyzed. The lowest furosine values were observed in artisanal hard-pressed cheeses (4.8 to 10.2 mg per 100 g of protein) and ripened with molds cheeses (4.2 to 12.8 mg per 100 g of protein). Industrial hard-pressed cheeses showed furosine values between 3.5 and 43.8 mg per 100 g of protein. Differences between samples may be attributed to the heat-treatment intensity given to cheese milk and to cheesemaking conditions. The highest contents of furosine were observed in processed cheeses (20 to 366.6 mg per 100 g of protein). A wide range of furosine content was found in fresh cheeses (17.9 to 73.6 mg per 100 g of protein), which could be due to the different amounts of dried milk added during the manufacture of cheeses.     

酿造原料及啤酒中镰刀菌毒素的研究进展

镰刀菌毒素是镰刀菌产生的次级代谢产物,能够通过感染酿造原料带入到啤酒中,从而时啤酒品质和人的健康造成威胁.为此,各国都制定了标准的检测方法和严格的限量标准.本文总结阐述了国内外有关啤酒酿造原辅料和成品啤酒中存在的镰刀菌毒素的性质、感染情况和检测方法等方面的研究进展,并对其在啤酒中的研究应用前景进行了展望.     

Contents of Fusarium toxins in Finnish and imported grains and feeds

The aim of the present study was to investigate the levels of trichothecenes in representative samples of Finnish and imported cereals and feeds from the 1987 and 1988 crops. Trichothecene concentrations were determined in samples as trimethylsilylether derivatives by a gas chromatograph equipped with a mass selective detector (GC-MSD) using selected ion monitoring (SIM) which proved to be a reliable and sufficiently sensitive technique. Representative samples of Finnish and imported oats, wheat, barley, rye, maize gluten, soy granules, rapeseed, turnip rapeseed, fish meal, poultry feed and pig feed were analysed employing 19-nortestosterone as an internal standard. Almost all grain and feed samples contained deoxynivalenol (DON) from 7 to 300 micrograms/kg and smaller amounts (13-120 micrograms/kg) of 3-acetyldeoxynivalenol (3-AcDON). The most toxic trichothecenes, T-2, HT-2 toxins and nivalenol (NIV) and also zeralenone (ZEN) were found at low concentrations in some samples. Six lots of oats containing 1.3-2.6 mg/kg of DON and 0.2-0.6 mg/kg of 3-AcDON were found in the Finnish grain samples. Silo samples of almost all imported grains contained DON but at levels below 50 micrograms/kg. Imported grains contained no other trichothecenes. DON, 3-AcDON and ZEN were found in imported feedstuffs at low concentrations.     

Survey of polybrominated diphenyl ether levels in Spanish commercial foodstuffs

Concentrations of 15 BDEs flame retardants have been determined in a large variety of food samples purchased in different markets across Spain. This is the first time that BDEs 184, 191, 196, and 197; impurities from BDEs formulations; and/or degradation products of BDE 209, have been detected in foodstuffs. Values ranged from <0.01 to 2482 pg/g fresh weight. The highest total BDE concentrations were found in fish samples (median of 189, range of 24-880 pg/g f.w.), followed by oils (median of 119, range of 14.8-2958 pg/g f.w.), meats (median of 75.9, range of 6.82-2518 pg/g f.w.), shellfish (median of 75.7, range of 3.29-677 pg/g f.w.), eggs (median of 73.5, range of 12.8-557 pg/g f.w.), and dairy products (median of 66.1, range of 3.24-1588 pg/g f.w.). The total BDE values found in this study are consistent with research reported elsewhere. They are in the same range as those recently reported by other European and Asian studies and lower than those conducted in the U.S. BDE 47 was the predominant congener in fish, shellfish, dairy products (except butter), and meats, while BDE 209 was the predominant in oil and egg samples. The most remarkable findings in this study were the large contribution of the highest brominated BDEs (hepta- to deca-BDE), and principally BDE 209, to the total BDE concentration found in Spanish foods, except fish and shellfish, and the presence of BDE 184, 191, 196, and 197 in many of the samples. The calculated intake of 38.5 ng/day of BDEs was comparable to intake assessment from other UE countries.     

Updated survey of Fusarium species and toxins in Finnish cereal grains

The aim of the project was to produce updated information during 2005–14 on the Fusarium species found in Finnish cereal grains, and the toxins produced by them, as the last comprehensive survey study of Fusarium species and their toxins in Finland was carried out at the turn of the 1960s and the 1970s. Another aim was to use the latest molecular and chemical methods to investigate the occurrence and correlation of Fusarium species and their mycotoxins in Finland. The most common Fusarium species found in Finland in the FinMyco project 2005 and 2006 were F. avenaceum, F. culmorum, F. graminearum, F. poae, F. sporotrichioides and F. langsethiae. F. avenaceum was the most dominant species in barley, spring wheat and oat samples. The occurrence of F. culmorum and F. graminearum was high in oats and barley. Infection by Fusarium fungi was the lowest in winter cereal grains. The incidence of Fusarium species in 2005 was much higher than in 2006 due to weather conditions. F. langsethiae has become much more common in Finland since 2001. F. graminearum has also risen in the order of importance. A highly significant correlation was found between Fusarium graminearum DNA and deoxynivalenol (DON) levels in Finnish oats, barley and wheat. When comparing the FinMyco data in 2005–06 with the results of the Finnish safety monitoring programme for 2005–14, spring cereals were noted as being more susceptible to infection by Fusarium fungi and the formation of toxins. The contents of T-2 and HT-2 toxins and the frequency of exceptionally high DON concentrations all increased in Finland during 2005–14. Beauvericin (BEA), enniatins (ENNs) and moniliformin (MON) were also very common contaminants of Finnish grains in 2005–06. Climate change is leading to warmer weather, and this may indicate more changes in Finnish Fusarium mycobiota and toxin contents and profiles in the near future.     

A critical review of biological methods for the detection of fungal toxins in foods and foodstuffs

Many biological assays (bioassays) have been developed to detect fungal toxins (mycotoxins). Few of these assays are as sensitive to mycotoxins as chemical assays or have been shown to be able to detect a wide range of different mycotoxins. Bioassays also suffer from a number of other problems including interference by nonfungal agents and from often being slow and less reproducible than chemical assays for mycotoxins. Bioassays have given little valuable information in their use in the surveillance of food and foodstuffs. Their major value has been in the initial identification and isolation of mycotoxins.     

Survey of dioxins, dioxin-like PCBs and marker PCBs in German feeds of plant origin

Within a representative survey the levels of dioxins (PCDD/PCDF), dioxin-like PCBs and marker PCBs in 206 German feed samples were analysed in the years 2004/2005. The sampling plan included compound feed (N = 115) and roughage and succulent feed (N = 91) and reflected the representative feeding situation in Germany. The median content of WHO-PCB-TEQ in analysed feed samples was 0.017 ng/kg [88% dry matter (d.m.)] and consequently more than a factor of 10 below the action level of 0.35 ng/kg (88% d.m.). A differentiation between compound feed and roughage and succulent feed showed that compound feed [median 0.007 ng/kg (88% d.m.)] were significantly lower contaminated with dioxin-like PCBs than roughage and succulent feed [median 0.058 ng/kg (88% d.m.)]. The median sum contents of the six marker PCBs were 0.16 μg/kg (88% d.m.) for compound feed and 0.56 μg/kg (88% d.m.) for roughage and succulent feed. The median of the WHO-PCDD/F-TEQ was 0.03 ng/kg (88% d.m.), the maximum level of 0.75 ng/kg (88% d.m.) was not exceeded. The median of the WHO-PCDD/F-PCB-TEQ was 0.05 ng/kg (88% d.m.) and consequently a factor of 25 below the maximum level of 1.25 ng/kg. Additionally the samples of roughage and succulent feed were analysed according to their contents of ash insoluble in HCl, representing a degree of the proportion of earthy components in feed. A slight correlation was found between ash insoluble in HCl and WHO-PCDD/F-TEQ (R ² = 0.59) whereas no correlation was found between ash insoluble in HCl and WHO-PCB-TEQ (R ² = 0.06).     

Survey of dioxins, dioxin-like PCBs and marker PCBs in German feeds of plant origin

Within a representative survey the levels of dioxins (PCDD/PCDF), dioxin-like PCBs and marker PCBs in 206 German feed samples were analysed in the years 2004/2005. The sampling plan included compound feed (N = 115) and roughage and succulent feed (N = 91) and reflected the representative feeding situation in Germany. The median content of WHO-PCB-TEQ in analysed feed samples was 0.017 ng/kg [88% dry matter (d.m.)] and consequently more than a factor of 10 below the action level of 0.35 ng/kg (88% d.m.). A differentiation between compound feed and roughage and succulent feed showed that compound feed [median 0.007 ng/kg (88% d.m.)] were significantly lower contaminated with dioxin-like PCBs than roughage and succulent feed [median 0.058 ng/kg (88% d.m.)]. The median sum contents of the six marker PCBs were 0.16 μg/kg (88% d.m.) for compound feed and 0.56 μg/kg (88% d.m.) for roughage and succulent feed. The median of the WHO-PCDD/F-TEQ was 0.03 ng/kg (88% d.m.), the maximum level of 0.75 ng/kg (88% d.m.) was not exceeded. The median of the WHO-PCDD/F-PCB-TEQ was 0.05 ng/kg (88% d.m.) and consequently a factor of 25 below the maximum level of 1.25 ng/kg. Additionally the samples of roughage and succulent feed were analysed according to their contents of ash insoluble in HCl, representing a degree of the proportion of earthy components in feed. A slight correlation was found between ash insoluble in HCl and WHO-PCDD/F-TEQ (R ² = 0.59) whereas no correlation was found between ash insoluble in HCl and WHO-PCB-TEQ (R ² = 0.06).     

Climatic models to predict occurrence of Fusarium toxins in wheat and maize

Although forecasting Fusarium infections have useful implications, it may be argued that forecasting Fusarium toxins is more useful to help reduce their entry into the food chain. Several disease incidence models have been commercialized for wheat, but only one toxin prediction model from Ontario, Canada, "DONcast", has been validated extensively and commercialized to date for wheat, and another has been proposed for maize. In the development of these predictive tools, the variation in toxin levels associated with year and agronomic effects was estimated from simple linear models using wheat and maize samples taken from farm fields. In wheat, environment effects accounted for 48% of the variation in deoxynivalenol (DON) across all fields, followed by variety (27%), and previous crop (14 to 28%). In maize, hybrid accounted for 25% of the variation of either DON or fumonisin, followed by environment (12%), and when combined 42% of the variability was accounted for. The robust site-specific, DON forecast model accounted for up to 80% of the variation in DON, and has been used commercially for 5 years in Canada. Forecasting DON and fumonisins in maize is more difficult, because of its greater exposure to infection, the role of wounding in infection, the more important role of hybrid susceptibility, and the vast array of uncharacterized hybrids available in the marketplace. Nevertheless, using data collected from controlled experiments conducted in Argentina and the Philippines, a model was developed to predict fumonisin concentration using insect damage and weather variables, accounting for 82% of the variability of fumonisins. Using mycotoxins as a measure of disease outcome, as opposed to disease symptoms, offers a more robust prediction of mycotoxin risk, and it accounts for mycotoxin accumulation that occurs frequently in the absence of any change in Fusarium symptoms.     

Removal of common Fusarium toxins in vitro by strains of Lactobacillus and Propionibacterium

This study was conducted to examine the ability of selected strains of Lactobacillus and Propionibacterium to remove common Fusarium toxins, trichothecenes, from liquid media. The trichothecenes studied were deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), nivalenol (NIV), fusarenon (FX), diacetoxyscirpenol (DAS), T-2 toxin (T-2) and HT-2 toxin (HT-2). The Lactobacillus rhamnosus strain GG (LGG), Lactobacillus rhamnosus strain LC-705 (LC-705) and Propionibacterium freudenreichii ssp. shermanii JS (PJS) were incubated in PBS buffer containing 20 µg toxin ml -1 for 1h at 37°C, and after centrifugation the concentration of the toxins was measured in the supernatant fraction. Both viable and heat-killed forms of LGG and PJS were more efficient than LC-705 in removing the toxins from the liquid media. LGG and PJS removed four of the seven tested toxins (the removal varying from 18 to 93%) and LC-705 two toxins (10-64%). Of the toxins, 3-AcDON was not removed by any of the bacteria; HT-2 was removed by the non-viable LGG and also slightly by non-viable LC-705; DAS was removed by all three bacteria tested. Binding is postulated as the possible mechanism of the removal, since no difference was observed between the ability of viable and heat-killed bacteria in removing the trichothecenes, and no degradation products of the toxins were detected by gas chromatography (GC)-mass spectrometry (MS) analysis. It is concluded that significant differences exist in the ability of the bacteria to bind trichothecenes in vitro.