Investigations into the occurrence of alkaloids in ergot and single sclerotia from the 2007 and 2008 harvests

As a contribution to the occurrence of ergot alkaloids in ergot from German rye and triticale, samples from the 2007 and 2008 harvests were analyzed. Twelve alkaloids—six pairs of main alkaloids and their corresponding epimers—were determined in extracts prepared under alkaline conditions by HPLC with fluorescence detection without preceding purification. The total alkaloid content was found to be 0.03–0.18% in ergot from rye (n = 19) and 0.06–0.22% in ergot from triticale (n = 4), respectively. Furthermore, single sclerotia (n = 40) were investigated in terms of alkaloid content and distributional pattern. The main alkaloids in ergot were ergocristine, ergotamine and ergocornine, although the alkaloid composition was highly variable. Presented in part at the 30th Mykotoxin-Workshop, Utrecht, The Netherlands, April 28–30, 2008     

Ergot alkaloids: Quantitation and recognition challenges

Bread, flour, infant formula and baby food samples (n=109, from which n=54 made of or containing rye), collected in 2001, 2003, and 2005, were analysed for ergot alkaloids. Samples were extracted using acidic conditions and the extracts subjected to an automated solid-phase clean up using combined cation exchange/reversed-phase sorbent cartridges (Oasis-MCX). Subsequent chromatographic separation and analysis was performed by liquid chromatography (LC) with fluorescence detection (FLD) and by LC with mass spectrometric detection (MS/MS). The ergot alkaloid (EAs) content of a sample was defined as the sum of the 16 alkaloids ergometrin(in)e, ergosin(in)e, ergotamin(in)e, ergostin(in)e, ergocornin(in)e, α-ergocryptin(in)e, β-ergocryptin(in)e and ergocristin(in)e. Comparability of results obtained by LC-FLD and LC-MS/MS was satisfactory, but varied for different alkaloids. The use of dihydro-ergocristine as an internal standard considerably improved the reliability of analytical data from LC-MS/MS. Compared with earlier data (Baumannet al., 1985) for median levels of ergot alkaloids in rye flour (140 ng/g) and bread (21.3 ng/g) from Switzerland, the median values for ergot alkaloids in rye flour collected in 2001 (n=13) and in 2005 (n=2) were 172 ng/g and 160 ng/g, respectively. The median values for bread (fresh weight) collected in 2001 (n=14), 2003 (n=7), and 2005 (n=2) were 87 ng/g, 120 ng/g, and 156 ng/g, respectively. Low levels of ergot alkaloids were also found in wheat products and in some infant formulae and baby foods containing rye. By additional LC-MS/MS experiments, the possible natural occurrence of ergot congeners containing the 9,10-unsaturated ergoline cation (m/z=223) was investigated. In a few samples, ergovalin(in)e was tentatively identified by these means. Presented at the 29^th Mykotoxin-Workshop, Fellbach, Germany, May 14–16, 2007     

Development of a UHPLC-FLD method for the analysis of ergot alkaloids and application to different types of cereals from Luxembourg

Ergot alkaloids are toxins produced by some species of fungi in the genus Claviceps, that may infect rye and triticale and, in a minor degree, other types of cereals. In this study, a new UHPLC-FLD method for the quantification of the six major ergot alkaloids as well as their corresponding epimers was developed. The sample preparation was done by a solid-liquid extraction with acetonitrile and clean-up via freeze-out. The method was fully validated and then applied to 39 samples (wheat, rye, triticale, and barley) harvested in Luxembourg in 2016. Samples were sieved (1.9?×?20 mm) prior to analysis in order to remove sclerotia, hosting the alkaloids. However, 23 samples still contained at least one ergot alkaloid >?LOQ and concentrations of the sum of the 6 ergot alkaloids ranged from 0.3 to 2530.1 μg/kg. Interestingly, the highest concentrations were measured in wheat and not in rye or triticale, suggesting that all kinds of cereals should be included in monitoring programs. The outcome of this study allowed giving a first overview of ergot alkaloid concentrations in cereals harvested in Luxembourg, and the measured concentrations were in similar ranges than in other parts of the world (e.g., Canada, France, Germany).     

Effects of the level of feed intake and ergot contaminated concentrate on ruminal fermentation and on physiological parameters in cows

The aim of the present study was to examine the effects of ergot contaminated feed concentrate at differing levels of feed intake on ruminal fermentation, and on various physiological parameters of dairy cows. Twelve double fistulated (in the rumen and the proximal duodenum) Holstein Friesian cows were fed either a control diet (on a dry matter (DM) base: 60% maize silage, 40% concentrate) or a diet containing ergot alkaloids (concentrate contained 2.25% ergot resulting in an ergot alkaloid concentration of the daily ration between 505 and 620 (μg/kg DM) over a period of four weeks. Daily feed amounts were adjusted to the current performance which resulted in a dry matter intake (DMI) variation between 6.0 and 18.5 kg/day. The resulting ergot alkaloid intake varied between 4.1 and 16.3 (μg/kg body weight when the ergot contaminated concentrate was fed. Concentrations of isovalerate, propionate and ammonia nitrogen in the rumen fluid were significantly influenced by ergot feeding, and the amount of ruminally undegraded protein, as well as the fermentation of neutral detergent fibre, tended to increase with the ergot supplementation at higher levels of feed intake, which might indicate a shift in the microbial population. Other parameters of ruminal fermentation such as ruminai pH, fermented organic matter as a percentage of intake, or the amount of non-ammonia nitrogen measured at the duodenum were not significantly influenced by ergot feeding. The activities of liver enzymes (aspartate aminotransferase, γ-glutamyltransferase, glutamate dehydrogenase, creatine kinase) in the serum were not affected by ergot feeding. The rectal measured body temperature of the cows significantly increased after ergot administration (p=0.019). Thus, body temperature can be regarded as a sensitive parameter to indicate ergot exposure of dairy cows.     

Natural occurrence of zeralenone in feeds and feedstuffs used in poultry and pig nutrition in colombia

A total of 200 samples of feedstuffs and mixed feeds used for poultry and pig nutrition in Colombia were analysed for zearalenone using a LC technique with a limit of detection of 20 μg/kg. Samples of grain sorghum, maize, processed soybean, rice meal, cottonseed meal, and poultry and pig feeds, representative of the Colombian production for the 1995–1996 harvest, were taken at feed manufacturing plants located in different cities of the country. Zearalenone was detected in 25 of 45 samples of sorghum, 2 of 33 samples of maize, 7 of 22 samples of rice meal, 9 of 17 samples of cottonseed meal, 11 of 30 samples of poultry feed and 6 of 16 samples of pig feed. Zearalenone was not detected in soybean or other feedstuff s analysed (wheat by- products, cassava meal, palm). Overall levels of zearalenone ranged from 29 to 3956 μg/kg, with a mean value of 436 μg/kg. Only one of the 6 positive samples of pig feed had a zearalenone concentration above 500 μg/kg, which is normally considered as the safe level for pigs.     

Mycotoxins in horse feed

A total of 62 samples of commercial horse feed preparations (complementary feeds) containing cereal mixtures (“muesli” or mash, n = 39; pelleted feeds, n = 12), and plain horse feed grains (maize, n = 5; oats, n = 4; barley, n = 2) were purchased from 21 different producers/distributors from the German market. All samples were analysed by competitive enzyme immunoassays (EIA) for six different mycotoxins (mycotoxin groups). Analytes (detection limit, mean recovery) were: deoxynivalenol (DON, 10 μg/kg, 84%), zearalenone (ZEA, 5 μg/kg, 93%), fumonisin B₁ (FB₁, 2 μg/kg, 113%), T-2 toxin (T-2, 0.1 μg/kg, 71%), sum of T-2 + HT-2 toxin (T-2/HT2, 0.2 μg/kg, 97%), ochratoxin A (OTA, 0.2 μg/kg, 67%), and total ergot alkaloids (Generic Ergot Alkaloids “GEA”, 30 μg/kg, 132%). All samples contained DON (16–4,900 μg/kg, median 220 μg/kg), T-2/HT-2 (0.8–230 μg/kg, median 24 μg/kg), and T-2 (0.3–91 μg/kg, median 7 μg/kg). ZEA was detected in 98% of the samples (7–310 μg/kg, median 61 μg/kg). Most samples (94%) were positive for FB₁ (2–2,200 μg/kg, median 27 μg/kg). Ergot alkaloids were detected in 61% of samples (28–1,200 μg/kg, median 97 μg/kg), OTA was found in 42% of samples (0.2–4 μg/kg, median 0.35 μg/kg). The results demonstrate that a co-contamination with several mycotoxins is very common in commercial horse feed from the German market. The toxin concentrations were in most cases well below the levels which are usually considered as critical or even toxic. The highest mycotoxin concentrations were mostly found in single-grain cereal feed: the maximum values for DON and FB₁ were found in maize, the highest T-2/HT-2 toxin concentrations were found in oats, and the highest concentration of ergot alkaloids was found in barley. In composed feeds, no correlation between cereal composition and mycotoxin levels could be found.     

Determination of ergot alkaloids in rye and rye flour

An effective and timesaving analytical method was developed for the determination of 12 ergot alkaloids (ergometrine, ergotamine, ergocristine, α-ergokryptine, ergosine, ergocornine, and their respective -inine isomers) in rye and rye flour. Samples were extracted with dichloromethane/ethyl acetate/methanol/aqueous ammonia (25%) (50/25/5/1, v/v/v/v), and extracts were purified using a basic alumina column. The eluate was dried in the nitrogen stream and redissolved in acetonitrile/ ammonia carbamate-buffer (0.2 g/1), (1/1, v/v), and injected into an HPLC-FLD system (λ_Ex 330 nm, λ_Em 415 nm), using the same mixture as mobile phase and a Phenyl-Hexyl column. Detection limits for the individual compounds ranged from 0.01 μg/kg to 0.5 μg/kg. In sample material spiked with a mixture of these compounds at two different levels (13 μg/kg and 27 μg/kg per compound), mean (n=5) recoveries were at 101% (s_r 6.4%) and 89% (s_r 3.1%), respectively. Presented at the 28th Mykotoxin-Workshop, Bydgoszcz, Poland, May 29–31, 2006     

Zum Einfluss einer hydrothermischen Behandlung auf den Ergotalkaloidgehalt von mutterkornbelastetem Roggen

Hydrothermal treatments are primarily used to increase the digestibility of nutrients and therefore to improve the feeding value of feedstuffs mainly for non-ruminants. Other positive side effects may occur, e.g. a decrease in toxicity of feed contaminated with mycotoxins. To study such effects, 4 batches of rye containing different percentages (0.8, 4.2, 8.3 and 25%) of ergot (Claviceps purpurea) were expanded and ergot alkaloid contents were analysed. After pre-conditioning of each batch by steam exposure for approx. 2 min, at 95 °C and 17% moisture, the material was expanded for approx. 5 sec. at 120 °C, 18% moisture, 40 bar mechanical pressure and 20 kWh/t mechanical energy input. Samples were collected before and after pre-conditioning and after expanding. Ergot alkaloids were analysed by HPLC. Analysis includedErgometrine, Ergotamine, Ergocornine, Ergocryptine, Ergocristine, Ergosine and their respective-inine isomers, the sum of these 12 ergot alkaloids was referred to as the total alkaloid content. On average, the hydrothermal treatment (pre-conditioning and expanding) caused a decrease of the total ergot alkaloid content of approx. 10%. Except for the batch containing 0.8% ergot, the efficiency of the hydrothermal treatment decreased with increasing ergot concentration in the batches. In general, the hydrothermal treatment changed the proportions of the ergot alkaloid isomers since the percentages of the-inine isomers of the total ergot alkaloid contents were increased with reduced-ine percentages. Whether this alteration is of toxicological relevance should be evaluated in animal experiments.     

Development of a method for the determination of citrinin in barley,rye and wheat by solid phase extraction on aminopropyl columns and HPLC-FLD

A method for the determination of the mycotoxin citrinin (CT) in rye, wheat and barley is described. The proposed method is based on ethyl acetate extraction, solid phase clean-up (SPE) on aminopropyl columns and reversed phase high performance liquid chromatography with fluorescence detection (RP-HPLC-FLD). The limits of detection and quantification of CT amounted to 0.6–0.9 μg/kg and 1.7– 3.3 μg/kg with mean recovery rates in the range of 77–92% (RSD 4.8–5.5%). This method can also be used for the determination of CT in red-fermented rice. Presented at the 29^th Mykotoxin-Workshop, Fellbach, Germany, May 14–16, 2007     

Mycotoxins in foods in Lower Saxony (Germany): results of official control analyses performed in 2009

In this presentation, the mycotoxin levels—as analysed by the analytical centre for mycotoxin surveillance of the state food laboratory (LAVES Braunschweig)—for approximately 500 food samples are reported. The samples were collected in the year 2009 at retail in the German federal state of Lower Saxony. Aflatoxin and ochratoxin A were analysed in dried fruits, spices, cereals and tree nuts. Ochratoxin A was detected in all samples of dried vine fruits, at levels up to 8.1 μg/kg. Aflatoxins and ochratoxin A were also found in nutmeg and curry powder: the maximum regulatory levels for aflatoxins were exceeded in 25% of the nutmeg samples. Nearly all samples of basmati rice contained aflatoxins, although at levels below the maximum regulatory level in all but one sample. Aflatoxins were also detected in about 50% of hazelnut samples, in 20% of the samples the maximum levels was exceeded (maximum 23.2 μg/kg). In contrast, aflatoxin contents in pistachios were surprisingly low. Fusarium toxins were analysed in cereals and cereal products such as flour, bread, and pasta. Deoxynivalenol (DON) was the predominant toxin found in these samples: DON was found in about 40% of the samples, although the maximum levels were not exceeded (max. 418 μg/kg). Fumonisins (FBs) and zearalenone (ZEA) were specifically analysed in maize products (snacks, flour and oil). Most of these samples (80%) were positive, but at levels not exceeding the maximum levels. Maximum levels were 98 μg/kg (ZEA) and 577 μg/kg (sum of FB₁ and FB₂). Ergot alkaloids (six major alkaloids) were analysed in rye flour, and approximately 50% were positive. The highest concentration of ergot alkaloids was 1,063 μg/kg; the predominant alkaloids were ergotamine and ergocristine. In conclusion, the results indicate that continuous and efficient control measures for mycotoxins in a wide range of critical foods are necessary to ensure compliance with maximum levels. Although the mycotoxin levels in the vast majority of samples were below maximum levels, year-to-year variation and changes in the production of relevant commodities may result in a different picture in the future.     

Analysis of rye grains and rye meals for ergot alkaloids

Due to the exceptionally hot and dry summer in 2003 the ergot of that harvest was rather small and could only be separated from normal grain with increased efforts. Based on a clean-up procedure of Wolffet al. (1) and of Kluget al. (2), a HPLC-FLD-method for the determination of 12 ergot alkaloids (6 “In”-, 6 “Inin”-forms) was established and modified. Actually reference substances are commercially available only for 5 selected alkaloids. Because of the instability of the alkaloids a new standard preparation procedure was tested and implemented. The maximum allowed impurity with ergot (0.05%=1000 μg alkaloids/kg) was exceeded in samples of harvest 2003. Except for one sample, all exceedings were detected in conventionally grown products, unlike organically grown products. Presented at the 27^th Mykotoxin-Workshop, Dortmund, Germany, June 13–15, 2005     

Co-occurrence of mycotoxins in swine feed produced in Portugal

A total of 404 samples of commercial swine feed from Portugal feed mills were analysed by HPLC methods for the presence of mycotoxins: 277 samples of feed for fattening pigs were analysed for ochratoxin A (OTA), zearalenone (ZEA), and deoxynivalenol (DON), and 127 samples of feed for sows were analysed for ZEA and fumonisins (FB₁ + FB₂). Concerning feed for fattening pigs, 21 (7.6%) samples were positive for OTA, (2–6.8 μg/kg), 69 (24.9%) were positive for ZEA (5–73 μg/kg), and 47 (16.9%) were positive for DON (100–864 μg/kg). In feed for sows, the results showed 29.9% of positive samples for ZEA (5–57.7 μg/kg) and 8.7% positive samples for FB₁ and FB₂ (50–391.4 μg/kg). Co-occurrence of DON/ZEA was found most frequently, but simultaneous contamination with OTA/ZEA and OTA/DON was also found.     

Occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in year 2003

The aim of this study was a monitoring of the occurrence ofAlternaria andFusarium mycotoxins in winter wheat from domestic crop in the year 2003. Altenuene was determined in 56 (100%) samples of winter wheat, range 14.5–41 μg/kg, mean 25 μg/kg. Alternariol was determined in 16 (28.6%) samples of winter wheat, range 6.3–22.1 μg/kg, mean 5.7 μ/kg. DON was determined in 42 (100%) samples of winter wheat, range 250–3500 μg/kg, mean 330 μg/kg. T2-toxin was determined in 42 (100%) samples of winter wheat, range 25–337 μg/kg, mean 99 μg/kg. ZEA was not determined in samples of winter wheat. Presented at the 26^th Mykotoxin-Workshop in Herrsching, Germary, May 17–19, 2004 Financial support. Supported (one part of experiments, the determination of Fusarium mycotoxins) by the Ministry of Agricu ture of the Czech Rebublic (Propect No QF3121)     

Dietary ergot alkaloids as a possible cause of tail necrosis in rabbits

This study describes the association between tail necrosis in rabbits and mycotoxins in rabbit feed. Clinical cases of tail necrosis were observed in 14 out of 103 rabbits kept in an outdoor group housing, fed with hay and a commercial pelleted feed. The observed clinical symptoms, alopecia, erosions, crusts and necrosis were restricted to the tail area and exclusively occurred in young rabbits aged 113?±?20 days. Dermatological examination suggested that ischemia had caused necrosis. Analysis of blood samples showed an elevated level of creatine kinase. No weight loss occurred in affected rabbits. Trauma caused by injuries or technopathic lesions was also excluded. Histopathologically, the lesions were characterized by acute muscle fibre degeneration and chronic active dermatitis with granulation tissue formation. Necropsy of one rabbit revealed hepatocellular degeneration and necrosis as remarkable findings. Feed analysis for ergot alkaloids by enzyme immunoassays yielded a mean and maximum ergot alkaloid content of 410?±?250 μg/kg and 1,700 μg/kg, respectively. Faeces of affected rabbits contained ergot alkaloids at levels up to 200 μg/kg. The mean and maximum dietary intake of total ergot alkaloids were 17 and 71 μg/kg bodyweight, respectively. Fusarium toxins (trichothecenes, zearalenone, fumonisins) were also found in the feed, but at levels which did not explain the observed effects. The results indicate that ergot alkaloids may have been the cause of tail necrosis, which is supported by literature data showing that rabbits are especially sensitive towards these toxins.     

Effects of different levels of ergot in concentrates on the growing and slaughtering performance of bulls and on carry-over into edible tissue

Abstract

The aim of the present study was to examine long-term effects of low levels of ergot alkaloids on growing bulls. Natural grown ergot with a mean total alkaloid concentrations of 633 mg/kg, and ergotamine (25%), ergocristine (15%) and ergosine (13%) as the most prominent alkaloids, was used. In a dose-response study 38 Holstein Friesian bulls were fed with three different doses of this ergot (0, 0.45 and 2.25 g/kg concentrate corresponding to an average total alkaloid concentration of the daily ration of 0, 69 and 421 µg/kg DM) over a period of approximately 230 days. Live weight, feed intake and health condition were monitored over the entire test period. The bulls were slaughtered at a live weight of approximately 550 kg. Carcass composition and quality were recorded and samples of liver, muscle, kidneys, fat, bile, urine and blood were analysed for ergot alkaloids. Liver enzyme activities and total bilirubin were measured in the blood. Statistically, no significant differences were detectable between the three feeding groups. Mean live weight gain over all groups was 1.41 kg/d with a mean dry matter intake of 7.35 kg/d. No carry over into tissues could be proved out of the experiment. To derive a no-effect level for beef cattle further research including higher ergot doses will be necessary.     

Interaction of benzo[c]phenanthridine and protoberberine alkaloids with animal and yeast cells

We compared the effects of four quaternary benzo[c]phenanthridine alkaloids – chelerythrine, chelilutine, sanguinarine, and sanguilutine – and two quaternary protoberberine alkaloids – berberine and coptisine – on the human cell line HeLa (cervix carcinoma cells) and the yeastsSaccharomyces cerevisiae andSchizosaccharomyces japonicus var. versatilis. The ability of alkaloids to display primary fluorescence, allowed us to record their dynamics and localization in cells. Cytotoxic, anti-microtubular, and anti-actin effects in living cells were studied. In the yeasts, neither microtubules nor cell growth was seriously affected even at the alkaloid concentration of 100 μg/ml. The HeLa cells, however, responded to the toxic effect of alkaloids at concentrations ranging from 1 to 50 μg/ml. IC₅₀ values for individual alkaloids were: sanguinarine IC₅₀ = 0.8 μg/ml, sanguilutine IC₅₀ = 8.3 μg/ml, chelerythrine IC₅₀ = 6.2 μg/ml, chelilutine IC₅₀ = 5.2 μg/ml, coptisine IC₅₀ = 2.6 μg/ml and berberine IC₅₀ >10.0 μg/ml. In living cells, sanguinarine produced a decrease in microtubule numbers, particularly at the cell periphery, at a concentration of 0.1 μg/ml. The other alkaloids showed a similar effect but at higher concentrations (5–50 μg/ml). The strongest effects of sanguinarine were explained as a consequence of its easy penetration through the cell membrane owing to nonpolar pseudobase formation and to a high degree of molecular planarity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods

In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 103 cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 103 but less than 104 cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 102 cfu/g sample to slightly more than 104 cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69–77.50 μg/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 μ};g/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25–252.50 μg/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00–289.38 μg/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25–436.25 μg/kg. Similarly, positive wheat flour samples were mostly collected from private homes. This revised version was published online in June 2006 with corrections to the Cover Date.     

The influence of ergot-contaminated feed on growth and slaughtering performance, nutrient digestibility and carry over of ergot alkaloids in growing-finishing pigs

Diets containing 0, 1 and 10 g ergot (Claviceps purpurea) per kg, corresponding to mean total alkaloid contents of 0.05, 0.60 and 4.66 mg/kg (sums of ergometrine, ergotamine, ergocornine, alpha-ergocryptine, ergocristine, ergosine and their -inine isomers analysed by a HPLC-method), were each fed ad libitum to 12 pigs in the BW range of 30-115 kg to study the effect of ergot-contaminated feed on growth and slaughtering performance and the carry over of ergot alkaloids. Additionally, balance trials were conducted to investigate the digestibility of nutrients. Tendencies towards reduced feed intake and BWG were observed at a feeding level of 4.66 mg total alkaloids per kg diet. Typical symptoms of ergot poisoning were not observed. Heart and spleen weights showed significant linear increases. Differences in carcass quality due to dietary treatment were not detected. No genuine ergot alkaloids were found in physiological samples. The balance trials demonstrated a significantly decreased protein digestibility for the most highly supplemented diet.     

T-2 and HT-2 toxin analysis in cereals and cereal products following IAC cleanup and determination via GC-ECD after derivatization

The authors present a new and sensitive method for the determination of T-2- und HT-2 Toxin in cereals and cereal products in the low ppb level. A representative part of the cereal sample is extracted with a mixture of methanol-water (90:10) and the extract is cleaned on the commercially available immunoaffinity column T-2test™ (IAC), eluted with methanol, derivatized by pentafluorpropionic anhydride (PFPA) and measured on a GC-ECD. The method has been successfully validated on wheat, rye and oats. The recovery rates with wheat and rye endowed on a level of 50 ppb and with 85 ppb naturally contaminated oats were 71–115% with a coefficient of variation of 5.7–19.5%. The detection limits of the method with a signal to noise level of 3:1 were 1.5–2.3 μg/kg for HT-2 and 1.1–1.7 μg/kg for T-2 toxin. Financial support: Federal Ministry of Food and Agriculture (part of the project 05HS 001 — Improvement and validation of type A trichothecene (T-2 toxin and HT-2 toxin) analysis and occurrence of these mycotoxins in food marketed in Germany)     

Effects of different levels of ergot in concentrates on the growing and slaughtering performance of bulls and on carry-over into edible tissue

The aim of the present study was to examine long-term effects of low levels of ergot alkaloids on growing bulls. Natural grown ergot with a mean total alkaloid concentrations of 633 mg/kg, and ergotamine (25%), ergocristine (15%) and ergosine (13%) as the most prominent alkaloids, was used. In a dose-response study 38 Holstein Friesian bulls were fed with three different doses of this ergot (0, 0.45 and 2.25 g/kg concentrate corresponding to an average total alkaloid concentration of the daily ration of 0, 69 and 421 microg/kg DM) over a period of approximately 230 days. Live weight, feed intake and health condition were monitored over the entire test period. The bulls were slaughtered at a live weight of approximately 550 kg. Carcass composition and quality were recorded and samples of liver, muscle, kidneys, fat, bile, urine and blood were analysed for ergot alkaloids. Liver enzyme activities and total bilirubin were measured in the blood. Statistically, no significant differences were detectable between the three feeding groups. Mean live weight gain over all groups was 1.41 kg/d with a mean dry matter intake of 7.35 kg/d. No carry over into tissues could be proved out of the experiment. To derive a no-effect level for beef cattle further research including higher ergot doses will be necessary.