Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1985

Trichothecene Mycotoxins Produced by Fusarium sporotrichioides Strain P-11 1

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A Visconti , CJ Mirocha , A Bottalico , and J Chelkowski

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Mycotoxin Research 1985, Vol. 1 (1), 3-10

A highly toxic strain of Fusarium sporotrichioides Sherb. (P-11) isolated from wheat in Poland produced on rice culture up to 11 trichothecenes, which are: T-2 toxin (750 ppm), neosolaniol (300 ppm), HT-2 toxin (75 ppm), acetyl T-2 toxin (35 ppm), 3´-hydroxy-T-2 (20 ppm), T-2 triol (12.5 ppm), 3´hydroxy-HT-2 (1.2 ppm), 4-acetoxy-T-2 tetraol (1.1 ppm), 15-acetoxy-T-2 tetraol (0.65 ppm), 8-acetoxy-T-2 tetraol (0.45 ppm), and T-2 tetraol (0.2 ppm). The presence of most of these trichothecenes, including the 3´-hydroxy-derivatives, in the excreta of animals treated with T-2 toxin indicates the existence of some correlation between T-2 toxin metabolism in animals and microorganisms, respectively. Addresses: 1, Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Via G. Amendola, 197/F, 70126 Bari, Italy; 2, Department of Plant Pathology, University of Minnesota, St. Paul, MN 55108, USA; 3, Department of Plant Pathology, Agricultural University, Warszawa 02-766, Poland.

Simultaneous Isolation of Xanthomegnin, Viomellein, Rubrosulphin, Viopurpurin, and Brevianamide A by Preparative HPLC CM Jansen and K Dose Mycotoxin Research 1985, Vol. 1 (1), 11-18

A method for the isolation of xanthomegnin, viomellein, rubrosulphin, viopurpurin, and brevianamide A from Penicillium viridicatum (DSM 2447) is described. After extraction, HPLC was performed with a preparative silicagel column, eluted with toluene/ethylacetate/formic acid (27/9/1, v/v/v) and dichloromethane/acetic acid (9/1, v/v). The toxins were detected with a UV-monitor. It was possible to isolate them in an absolutely pure state. The described method is operationally simple and very efficient. Address: Institut für Biochemie, Johannes-Gutenberg-Universität, Johann-Joachim-Becher-Weg 30, D-6500 Mainz, Germany.

Production of 3-Acetyldeoxynivalenol in Shake Culture K Ishii, H Sato, and Y Ueno Mycotoxin Research 1985, Vol. 1 (1), 19-24

In order to produce deoxynivalenol (DON), conditions for the production of 3-acetyldeoxynivalenol (Ac-DON) in shake culture were studied. A selected isolate of DON-producer, F. graminearum strain R 2118, produced about 70 mg/mL of Ac-DON when cultured at 25 °C for 4 days in a medium consisting of 3% sucrose, 0.1% peptone, and 0.1% yeast extract. From the mass culture (10 L) of the strain, 792 mg of Ac-DON were isolated and 384 mg of DON were obtained after hydrolysis of the acetate. Address: Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Shinjuku-ku, Tokyo 162, Japan.

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Optical Motility Test for the Detection of Trichothecenes Using Brine Shrimps R Schmidt Mycotoxin Research 1985, Vol. 1 (1), 25-29

A new optical bio-assay using brine shrimp larvae (Artemia salina L.) has been successfully applied for monitoring the presence of highly toxic trichothecenes. The motility of the swimming animals was determined instead of their lethality. This permits more objective and precise results. The inhibition of the motility can be seen earlier than the death of animals in toxicity tests. The relationship between time and dose is indicated for two different concentrations. This test can also be applied in the control of other harmful compounds. Address: Fa. Hans W. Schmidt, Saarstr. 52, D-6500 Mainz, Germany.

Chemical Analysis for Ochratoxin Poisoning HM Stahr1, M Domoto1, Bei Lei Zhu2, and R. Pfeiffer3 Mycotoxin Research 1985, Vol. 1 (1), 31-35

No abstract available. Addresses: 1, Chemistry Laboratory, Veterinary Diagnostic Laboratory, Iowa State University, Ames, Iowa 50011, U.S.A.; 2, Veterinary College, Beiping University, Beiping, China; 3, Toxicology Department, Veterinary College, University of Illinois, Urbana, Illinois, USA.

Patulin Degradation in Saccharomyces cerevisiae: Sensitive Mutants 1 1 2 P Thonart , ZL Sumbu , and J Bechet

Mycotoxin Research 1985, Vol. 1 (1), 37-40

No abstract available. Addresses : 1, Faculté des Sciences Agronomiques de l‘Etat, 5800 Gembloux, Belgique; 2, Institut de Recherches et Institut des Industries de Fermentation CERIA, 1070 Bruxelles, Belgium.

Inducing Effect of Testosterone on the Hepatic Reduction of Zearalenone in the Female Prepubertal Rat Monica Olsen Mycotoxin Research 1985, Vol. 1 (2), 51-56

The prepubertal responsiveness of 3-hydroxysteroid dehydrogenase and zearalenone-reducing activity to either a large subcutaneous dose of testosterone or dietary zearalenone was investigated in female rats. Testosterone induced both the activity of the 3-hydroxysteroid dehydrogenase, with androsterone as substrate, and zearalenone reduction to - and -zearalenol. Zearalenone had no effect on the activity of 3-hydroxysteroid dehydrogenase, but had a slight inducing effect on the zearalenone reduction to -zearalenol. Both testosterone and zearalenone had a growth-promoting effect on uterus. Address: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, S-75007 Uppsala, and the Department of Zoophysiology, University of Uppsala, Sweden.

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Immunotoxicity of Repeated Low Level Exposure to T-2 Toxin, a Trichothecene Mycotoxin, in CD-1 Mice MJ Taylor, RV Reddy, and RP Sharma Mycotoxin Research 1985, Vol. 1 (2), 57-64

Male CD-1 mice were gavaged with T-2 toxin (0.0–5.0 mg/kg body weight) every third day. Body weight gain was depressed by exposure to 2.5 mg/kg, or greater, T-2 toxin; this was not associated with decreased food intake. The weights of the liver, kidney, spleen, and thymus were affected by two weeks exposure to T-2 toxin. However, a persistent effect after four weeks was observed only for the thymus. Peripheral leucocyte counts were elevated in the highest dose groups after two and four weeks. Thymidine uptake by cells not simultaneously exposed to mitogen was increased in splenic cell cultures of mice exposed to 2.5 mg/kg T-2 toxin for two or four weeks. Phytohemagglutinin stimulation of splenic lymphocytes following two weeks of exposure was depressed in the 2.5 mg/kg dose group; this phenomenon was not observed after four weeks exposure. Response to pokeweed mitogen increased after four weeks of exposure to 2.5 mg/kg T-2 toxin. A delayed-type hypersensitivity response decreased following two weeks exposure to levels greater than 0.02 mg/kg. Production of IgM class antibodies by splenic lymphocytes, evaluated by a hemolytic plaque response to sheep erythrocytes, was depressed in the 2.5 mg/kg dose group after two weeks exposure to T-2 toxin. The sensitivity and specificity of T-2 toxin immunotoxicity was indicated by the various parameters evaluated. Address: Toxicology Program, Utah State University, Logan UT 84322-5600, USA. Effect of Different Antioxidants and Free Radical Scavengers on Aflatoxin Production C Fanelli1, AA Fabbri1, S Pieretti1, E Finotti2, and S Passi2 Mycotoxin Research 1985, Vol. 1 (2), 65-69

Different antioxidants and free radical scavengers on aflatoxin production are analysed. The different compounds at different concentration were used: buthylated hydroxyanisole (BHA), buthylated hydroxytoluene (BHT), -tocopherol (vitamin E), ascorbic acid (vitamin C), reduced glutathione, cysteine, cysteamine. The above compounds were tested in culture of Aspergillus parasiticus supplemented with carbon tetrachloride, a potent stimulating agent of aflatoxin biosynthesis. Cysteamine and BHA highly inhibited the aflatoxin production induced by carbon tetrachloride, the inhibition decreased by lowering the concentration. On the contrary, vitamin E, vitamin C, reduced glutathione and cysteine further enhanced the carbon tetrachloride stimulating effect. The addition of the above compounds did not significantly affect the growth of the fungal mycelia. Addresses: 1, Dipartimento di Biologia vegetale, Università di Roma ’La Sapienza’, Largo Cristina di Svezia, 2400165 Roma, Italy; 2, Istituto San Gallicano (IFO), Via San Gallicano 25a, 00165 Roma, Italy. Aflatoxin M1 in Milk in Southern Italy A Visconti, A Bottalico, M Solfrizzo Mycotoxin Research 1985, Vol. 1 (2), 71-75

A survey on the occurrence of aflatoxin Ml was carried out in the province of Bari (Italy) on a total of 106 samples of raw milk from local farms, commercially heat-treated milk, and dried milk. About 72 % of the samples were contaminated with aflatoxin Ml within the concentration range 4–280 ng/kg. The incidence of aflatoxin Ml was much higher in commercial milk (91 %) than in farm milk (26 %), and all the dried milk samples contained aflatoxin Ml (10 to 280 ng per kg of powder milk). Address: Istituto tossine e micotossine da parassiti vegetali, Consiglio Nazionale delle Ricerche, 70126 Bari, Italy. 175

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1985

Biotransformation of T-2 Toxin and Diacetoxyscirpenol in the Isolated Perfused Rat Liver M Gareis, B Ertl, J Bauer, and B Gedek Mycotoxin Research 1985, Vol. 1 (2), 77-82

The major metabolite found in the perfusate of liver perfused with 2 mg of T-2 toxin was HT-2 toxin. TLC-analysis revealed that a total of 0.29 mg of HT-2 toxin was excreted into the perfusate after 30 min of perfusion. The toxin was detected as early as at 3 min of perfusion and reached a maximum of concentration at 6 min of perfusion (40% of the total amount). Decreasing amounts were detected until the end of the experiment (30 min). Minor metabolites (less than 10% compared to HT-2 toxin) and non-transformed T-2 toxin were mainly found between 3 and 9 min of perfusion. By GLC-MS analysis, T-2 toxin and 12 metabolites were determined in the combined perfusate at 9 min of perfusion. Three as yet not described minor metabolites of T-2 toxin were shown to occur in the liver perfusate. Two of these compounds, whose mass spectra contained all of the characteristic ions of neosolaniol TFA were eluted at 6:24 min and 7:26 from the capillary column and are supposed to be isomers of neosolaniol. The two metabolites of diacetoxyscirpenol (DAS, 2 mg) in the perfusate of isolated rat liver were monoacetoxyscirpenol (MAS) and scirpenetriol (SCT). Non-transformed DAS (0.106 mg) and MAS (0.190 mg) were found up to 9 min of perfusion, while SCT was detected in perfusate samples from 6 to 30 min of perfusion. Address: Institute for Medical Microbiology, Infectious and Epidemic Diseases, Faculty of Veterinary Medicine, University of Munich, Veterinärstr. 13, 8000 Munich 22, Germany. Moniliformin; a 13C and 1H NMR Study John C Mitchell1, M John Perkins1 and John Gilbert2 Mycotoxin Research 1985, Vol. 1 (2), 83-86

The mycotoxin moniliformin has been shown to be produced by a number of Fusarium species under culture conditions, and has also been shown to co-occur naturally with deoxynivalenol and zearalenone in visibly mouldy corn from South Africa. In view of the widespread occurrence of Fusarium species in cereal crops, it was decided to extend previous surveillance of Fusarium mycotoxins in the United Kingdom to include moniliformin. As moniliformin reference material was not available commercially a quantity of moniliformin was synthesized according to the method described by Bellus et al. This note describes preliminary characterisation prior to development of suitable sensitive analytical methods. Addresses: 1, Chelsea College, Department of Chemistry, University of London, London SW3, United Kingdom; 2, Ministry of Agriculture, Fisheries and Food, Food Science Laboratory, Haldin House, Queen Street, Norwich NR2 4SX, United Kingdom.

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Etiology of Turkey “X“ Disease in Retrospect: A Case for the Involvement of Cyclopiazonic Acid Richard J Cole Mycotoxin Research 1986, Vol. 2 (1), 3-7

The etiology of the classical turkey “X“ disease syndrome is reappraised based on original reports in conjunction with current information. The clinical signs described in those original reports cannot be totally explained as being typical for aflatoxicosis. The unexplained effects of the disease can be resolved by the proposed presence of the mycotoxin cyclopiazonic acid which is frequently produced by Aspergillus flavus along with the aflatoxins. Address: USDA, ARS, National Peanut Research Laboratory, 1011 Forrester Drive, S. E., Dawson, Georgia 31742.

Isolation of Secondary Metabolites of Aspergillus ochraceus by HPLC Ivonne Delgadillo Mycotoxin Research 1986, Vol. 2 (1), 9-17

A method is described for the isolation and purification of ochratoxin A, ochratoxin B, ochratoxin ß, mellein, 4-hydroxymellein and penicillic acid produced by Aspergillus ochraceus in a synthetic liquid medium. Ochratoxin , which was not found in the culture medium, was obtained by acid hydrolysis of ochratoxin A. A high pressure liquid chromatograph equipped with Lichrosorb 100 and Lichrosorb RP-18 columns and UV and/or Refractive Index detectors was used. Address: Universidade de Aveiro, Departamento de Química, 3800 Aveiro, Portugal. The Isolation and Identification of Some Toxic Constituents of Aspergillus wentii Wehmer CJ Rabie 1, PS Steyn2 and FR van Heerden2 Mycotoxin Research 1986, Vol. 2 (1), 19-24

Extraction of a maize culture of a toxinogenic strain of A. wentii led to the isolation and characterization of three anthraquinones, three bianthrones, a xanthone and a benzophenone. The structures were derived from spectroscopic data and were supported by chemical degradation. Of these, emodin, 1,6di-0-methylemodin, 5-0-methylsulochrine and 1,3-di-0 -methylemodin bianthrone were mildly toxic to ducklings. Addresses: 1, National Research Institute for Nutritional Diseases, Medical Research Council, PO Box 70, Tygerberg, 7505, Republic of South Africa; 2, National Chemical Research Laboratory, Council for Scientific and Industrial Research, PO Box 395, Pretoria, 0001, Republic of South Africa. Isolation, Purification and Nuclear Magnetic Resonance Spectra of Pulvilloric Acid Jill Barber, Simon M Bradley and Sarah M Pope Mycotoxin Research 1986, Vol. 2 (1), 25-32

A strain of Penicillium pulvillorum which produces pulvilloric acid has been identified. Pulvilloric acid has been isolated and purified and modern techniques have been used to assign its 13C and 1H nuclear magnetic resonance (NMR) spectra. Address: Department of Pharmacy, University of Manchester, Manchester M 13 9 PL, United Kingdom. 177

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The Effects of Demethylsterigmatocystin and Sterigmatin on ATP Synthesis System in Mitochondria: A Comparison with Sterigmatocystin 1 2 2 2 3 Kiyoshi Kawai , Teruhiko Nakamaru , Kazuo Hisada , Yoshinori Nozawa , and Hideki Mori

Mycotoxin Research 1986, Vol. 2 (1), 33-38

The effects of demethylsterigmatocystin and its structural isomer, sterigmatin, on oxidative phosphorylation of mitochondria were studied by means of isolated rat liver mitochondria in comparison with sterigmatocystin. Both compounds were found to uncouple the oxidative phosphorylation in mitochondria, causing marked decreases in RC index and P/O ratio. Sterigmatin, in which dihydrobisfuran ring and xanthone nucleus are combined in a linear manner, was evidently more toxic to mitochondrial functions than demethylsterigmatocystin which was an angular molecular shape. Though their uncoupling concentrations were rather high for assessing their in vivo toxicity, a good correspond was observed between their orders of potencies in the toxicity to mitochondrial functions and in the cytotoxicity to rat hepatocytes. The demethylation of sterigmatocystin resulted in an obvious decrease of uncoupling activity. Addresses: 1, Department of Food and Nutrition, Faculty of Home Economics, Chukyo Women‘s University, Ohbu City 474, Aichi, Japan; 2, Department of Biochemistry and 3, Department of Pathology, Gifu University School of Medicine, Tsukasamachi 40, Gifu City 500, Japan.

HPLC of Trichothecenes - Separation of Neosolaniol, NT-1 Toxin, and NT-2 Toxin Rainer Schmidt Mycotoxin Research 1986, Vol. 2 (1), 39-43

A high pressure liquid chromatographic (HPLC) method is described for the isolation of trichothecenes formed on moldy rice. Extraction of the cultures was followed by purification and fractionation with a C18-Sep Pak cartridge. The polar fraction contained neosolaniol, 4,8-diacetoxy-12,13epoxytrichothec-9-ene-3,15-diol (NT-1) and 4-acetoxy-12,13-epoxytrichothec-9-ene-3,8,15-triol (NT2), while in another fraction HT-2 toxin, T-2 toxin and acetyl-T-2 toxin were eluted. A high pressure liquid chromatograph equipped with a C18 μBondapak column and an RI-detector (differential refractometer) were used for the isolation procedure. Address: Firma Hans W. Schmidt, Saarstr. 52, D-6500 Mainz 1, Germany.

Deoxynivalenol and 3-acetyldeoxynivalenol – Mycotoxins Associated with Wheat Head Fusariosis in Poland Angelo Visconti1, Jerzy Chelkowski2, and Antonio Bottalico1 Mycotoxin Research 1986, Vol. 2 (2), 59-64

Samples of wheat naturally infected in the field by Fusarium culmorum (W.G.Sm.) Sacc. and Fusarium graminearum Schwabe were analyzed for deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone. No zearalenone was detected at levels higher than 0.5 mg/kg. Deoxynivalenol was present in 100% and 3-acetyldeoxynivalenol in 80% of the examined samples at levels of 0.21 to 30.4 mg/kg and 0.54 to 29.54 mg/kg, respectively. The mycotoxin levels in the chaff were 5 to 50 times higher than in the kernels. This is the first report on natural occurrence of 3-acetyldeoxynivalenol in wheat. This toxin, in addition to deoxynivalenol, was highly correlated with wheat head fusariosis. These findings suggest that more attention should be given to the occurrence of 3-acetyldeoxynivalenol in cereal grains during the growth as well as during storage. Addresses: 1, Istituto Tossine e Micotossine da Parassiti Vegetali, C N R, 70126 Bari, Italy; 2, Department of Plant Pathology, Agricultural University, 02 -766 Warsaw, Poland. 178

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Simultaneous Quantitative Determination of Secondary Metabolites of Aspergillus ochraceus by TLC Ivonne Delgadillo Mycotoxin Research 1986, Vol. 2 (2), 65-69

A simple TLC method for the quantitative determination of mellein, 4-hydroxymellein, penicillic acid, ochratoxin A, B,  and  is described. Application of this technique permits metabolic studies of the influence of different factors on the formation of Aspergillus ochraceus metabolites. Address: Universidade de Aveiro, Departamento de Quimica, P-3800 Aveiro, Portugal.

Effects of Nivalenol on Pregnancy and Fetal Development of Mice Y Itol, K Ohtsubo2, K Ishii3, and Y Ueno3 Mycotoxin Research 1986, Vol. 2 (2), 71-77

Nivalenol (3,4,7,15-tetrahydroxy-12,13-epoxytrichothec-9-en-8-one) was injected intraperitoneally to pregnant mice at dose levels of 0, 0.1, 0.5 or 1.5 mg/kg b.w./day on days 7–15 of gestation. The highest dose caused stillbirths after vaginal hemorrhage in six of ten animals. High embryo lethality was recorded in the 2 highest dose groups (87.8 and 48.4%). No fetal malformations were observed in the test groups. A single administration of 3 mg/kg on day 7 affected the embryos within l0 hrs, damaged the placenta within 24 hrs, and revealed stillbirths at 48 hrs. Addresses: 1, Institute for Medical Science of Aging, Aichi Medical University, Nagakute-cho, Aichi-gun, Aichi, Japan; 2, Department of Clinical Pathology, Tokyo Metropolitan Institute of Gerontology, Sakae-cho, Itabashi-ku, Tokyo, Japan; 3, Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Science, Science University of Tokyo, Ichigaya funakawaramachi, Shinjuku-ku, Tokyo, Japan.

Cytotoxicity of Fusarium T-2 in Cultured Madin-Darby Bovine Kidney (MDBK) and Primary Fetal Bovine Kidney (PFBK) Cells Masao Yoneyama and Raghubir P Sharma Mycotoxin Research 1986, Vol. 2 (2), 79-88

Cytotoxicity of Fusarium T-2 toxin was evaluated in cultured Madin-Darby bovine kidney (MDBK) and primary fetal bovine kidney (PFBK) cells. The criteria for evaluation included number of adherent cells, phase contrast microscopy, and the scanning and transmission electron microscopy. The primary cells were more sensitive to the toxic effects of T-2 toxin than MDBK cultures. Cytotoxicity was observed when the cultures were exposed to the toxin for only 1 hour and then incubated with untreated media. Cell multiplication was decreased in both systems in 72 hour cultures. Scanning electron microscopy indicated loss of inter-cell contact and marked alterations in cell shape. Transmission electron microscopy indicated extensive proliferation of lysosomal bodies and proliferation of endoplasmic reticulum. The membrane system and mitochondria were not affected. Results indicated the kidney cells are highly sensitive to the toxic effects of T-2 toxin. Address: Toxicology Program, Utah State University, Logan, UT 84322-4620, USA.

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Studies on the Embryotoxicity and Mutagenicity of Mycotoxins G Matthiaschk and A Korte Mycotoxin Research 1986, Vol. 2 (2), 89-97

Embryotoxicity: Aflatoxin B1 (AFB1), G1 (AFG1), and Patulin (PA) were investigated in NMRI mice for embryotoxic and teratogenic activity. These three mycotoxins were injected intraperitoneally or given orally on day 12 and 13 of pregnancy. AFB1 (15, 45, and 90 mg/kg ip or 45 mg/kg po) produced a moderate retardation in the fetal development and a dose related increase of cleft palates, wavy ribs, and diaphragm changes. The effects after injection of AFG1 (45 and 90 mg/kg ip) were reduction of fetal weights, increase of diaphragm changes, and malformations of kidneys. PA (1.25, 2.5, and 3.75 mg/kg ip or 3.75 mg/kg po) elevated the rate of cleft palates after 3.75 mg/kg. In the dominant lethal assay neither PA (2.5 and 5 mg/kg ip) nor AFB1 (15 and 45 mg/kg ip) increased the frequency of the dominant lethal mutations. Both mycotoxins showed no mutagenic activity in this test system. The capability of AFB1, AFG1, and PA to induce chromosome damages in vivo was tested in the Chinese Hamster by examination of bone marrow cells, each after two oral doses (AFB1: 12.5 and 25 mg/kg; 25 and 50 mg/kg; PA: 10 and 20 mg/kg). The three mycotoxins induced chromosome aberrations in the following order of activity: PA>AFB1>AFG1. Address: Abteilung Toxikologie, Max von Pettenkofer-Institut, Bundesgesundheitsamt, Postfach 330013, D-1000 Berlin 33, Germany.

Production of Ochratoxin A, Citrinin, and Ergosterol By Penicillium viridicatum in Autoclaved and Non-Autoclaved Wheat at Low Temperature Angela Boley and Hans-Martin Müller Mycotoxin Research 1986, Vol. 2 (2), 99-103

Wheat (moisture content: 26 %) was autoclaved or left untreated, inoculated with conidia of Penicillium viridicatum and stored at 10 °C. The fungus grew on both substrates and was the dominant mould on the non-autoclaved grain. Autoclaving resulted in an earlier onset of ergosterol, ochratoxin A, and citrinin production due to accelerated mould growth. Yield of ochratoxin A increased while citrinin slightly decreased in autoclaved wheat. Address: Institute of Animal Nutrition, University of Hohenheim, D-7000 Stuttgart 70, Germany.

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Elimination of Roquefortine in the Rat I Laws and PG Mantle Mycotoxin Research 1987, Vol. 3 (1), 3-6 14

C-roquefortine, given to rats per os or intraperitoneally, was eliminated principally by the biliary route. Most of the given radiolabel accounted for had been voided within one day of administration. Roquefortine was not metabolised in rat liver homogenate but significant transformation to more polar products was evident in faeces. Address: Biochemistry Department, Imperial College, London SW7 2AZ, United Kingdom.

Chromatographic Quantitation of Territrems A, B, and C, Tremorgenic Mycotoxins from Aspergillus terreus KH Ling, MF Chang, SC Tsai, YW Peng, BJ Chen, and EC Wang Mycotoxin Research 1987, Vol. 3 (1), 7-12

This paper describes assay procedures for territrems A, B, and C by thin layer chromatography (TLC)fluorodensitometry and reverse phase high performance liquid chromatography (HPLC). Address: Institute of Biochemistry, College of Medicine, National Taiwan University, Jen-ai Road, No. 1, Section 1, Taipei, Taiwan, Republic of China.

Methods of Tracing Hydrogen in Polyketide Biosynthesis: High Field NMR Spectroscopy of Citrinin Produced in a D2O Based Medium 1 2 Jill Barber , Anne C Chapman , and Tina D Howard

Mycotoxin Research 1987, Vol. 3 (1), 13-18

Penicillium citrinum cultures have been germinated on an H2O-based medium, resuspended on a D2Obased medium and treated with [1,2-13C2] acetate. The resulting citrinin has been analysed by 2H and 13 C nuclear magnetic resonance spectroscopy and information about the metabolism of hydrogen in citrinin biosynthesis has been deduced. Addresses: 1, Department of Pharmacy, University of Manchester, Manchester, M13 9PL, UK; 2, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.

Prevention of Aflatoxin Contamination Through Some Commercial Chemical Products and Plant Extracts in Groundnut MP Ghewande and G Nagaraj Mycotoxin Research 1987, Vol. 3 (1), 19-24

The effect of rock salts, NaCl, propionic acid, NCP.75, plant products - asafoetida, turmeric powder and aqueous leaf extracts of Azadirachta indica, Lawsonia alba, Pongamia glabra and Tridax procumbens on seed colonization and aflatoxin production by Aspergillus flavus (NRRL-3000) was studied in two Spanish bunch groundnut varieties (J-11 and JL-24). All these treatments inhibited seed colonization and aflatoxin production to varying degrees. Inhibition of seed colonization with chemicals, plant products and aqueous leaf extracts was observed to range between 17 to 96 %, 27 to 100 %, and 8 to 75 % while inhibition of aflatoxin production ranged from 14 to 74 %, 42 to 71 %, and 6 to 64 %, respectively. In general, salts (20 g/L), propionic acid (10 mL/L), asafoetida (pure 1 g/L and

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impure 20 g/L), and Azadirachta indica aqueous leaf extract (20 g/L) are better in preventing aflatoxin contamination in both the groundnut varieties. Address: National Research Centre for Groundnut, Junagadh 362015, Gujarat, India.

On the Metabolism of the Mycotoxin Deoxynivalenol in the Isolated Perfused Rat Liver M Gareis, J Bauer, and B Gedek Mycotoxin Research 1987, Vol. 3 (1), 25-32

An isolated rat liver was perfused with deoxynivalenol (DON) at a dose of 3 mg in a recirculating perfusion system. To identify glucuronide conjugates equal amounts of bile samples, perfusate and liver homogenates were incubated with and without (control) a ß-glucuronidase preparation and analyzed by thin layer chromatography and capillary gas liquid chromatography - chemical ionization mass spectrometry. A total of 40.4 % of the administered dose of DON was found to be conjugated with glucuronic acid (perfusate 20.4 %, bile 19.2 %, liver 0.8 %), while only 1.3 % of the parent DON (perfusate 1.1 %, bile 0.2 %) was detected. The cleavage of DON-glucuronide was demonstrated by incubating DON-glucuronide containing bile samples with intestine contents under anaerobic conditions. Address: Institute for Medical Microbiology, Infectious and Epidemic Diseases, Faculty of Veterinary Medicine, University of Munich, Veterinärstrasse 13, D-8000 Munich 22, Germany.

A Rapid Extraction Method for Detecting Aflatoxin Producing Isolates a Miguel A Moreno, M del Carmen Ramos, and Guillermo Suàrez

Mycotoxin Research 1987, Vol. 3 (1), 33-35

A rapid extraction method for screening aflatoxin producing potential of Aspergillus flavus group isolates is described. The method is performed using a moist wheat medium with ca. five infected grains extracted with 2 mL of chloroform, and using thin layer chromatography. This method was proved with 95 A. flavus isolates from animal feeds. Address: Unidad Docente de Microbiologia, Departamento de Patologia Animal I (Sanidad Animal), Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.

Stability of Citrinin and Deoxynivalenol During Germination Process of Barley AA El-Banna Mycotoxin Research 1987, Vol. 3 (1), 37-41

The stability of citrinin and deoxynivalenol during germination process of barley spiked with these mycotoxins at a level of 2 μg/g was investigated. Germinated barley was analyzed after 1, 3, and 5 days to follow the stability of citrinin and deoxynivalenol during the germination process. Two thin layer chromatographic methods were used for determinations of citrinin and deoxynivalenol. An average of 93.6 % of citrinin and 77.1 % of deoxynivalenol were destroyed within 5 days during the germination process of barley. Address: Institute for Microbiology, Toxicology and Histology, Federal Centre for Meat Research, Kulmbach, Germany.

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Cleavage of Territrems by Alkaline Hydrogen Peroxide: Isolation and Characterization of the Aromatic Moiety of Territrems Kuo Huang Ling, Bai Jiun Chen, Yun Wen Peng, Shih Chong Tsai, Fu Chuo Peng, and Chung Kuang Yang Mycotoxin Research 1987, Vol. 3 (2), 58-64

The chemical reaction of cleavaging territrem B to give 3,4,5-trimethoxy benzoic acid by alkaline hydrogen peroxide was investigated. The method was applied for confirmation of the chemical structure of the aromatic moiety of territrem A, A´, B, and B´. The physicochemical properties of the aromatic cleavage product of territrem A indicated the structure as 3,4-methylendioxy, 5-methoxy benzoic acid (or 4-methoxy,6-carboxy,1,3-benzodioxole). The experiment also gave the evidences that territrem A and A´, on the other hand territrem B and B´ have the identical aromatic moieties on their structures. Address: Institute of Biochemistry, College of Medicine, National Taiwan University, Taipei, Taiwan, Republic of China.

Isolation of Macrocyclic and Non-Macrocyclic Trichothecenes (Stachybotrys and Fusarium Toxins) from the Environment of 200 Ill Sport Horses 1 2 3 3 Balázs Harrach , Arpád Bata , Gabriella Sándor , and András Ványi

Mycotoxin Research 1987, Vol. 3 (2), 65-68

Satratoxins H and G, verrucarin J, and roridin E were isolated from the bedding straw of 200 sport horses exhibiting typical symptoms of stachybotryotoxicosis. At the same time, the oat feed consumed by the horses contained non-macrocyclic Fusarium trichothecenes: T-2 toxin and diacetoxyscirpenol. Addresses: 1, Veterinary Medical Research Institute, Hungarian Academy of Sciences, H-1581 Budapest, POB 18, Hungary; 2, Department of Biochemistry and Food Technology, Technical University, Budapest, Hungary; 3, Central Veterinary Institute, Budapest, Hungary.

Toxicity of Epoxy Trichothecenes in Cultured Mammalian Cells KE von Milczewski Mycotoxin Research 1987, Vol. 3 (2), 69-76

With four established cell lines, cytotoxicity of 23 epoxy trichothecenes was tested. Three cell lines from human origin have been found to have a similar degree of sensitivity to trichothecenes. A pig kidney cell line had a higher sensitivity to NT-1 toxin, neosolaniol, tetraacetyl T-2, and iso-T-2 toxin. The potential cause for discrepancies in literature were discussed. Activity of esterases in calf serum used for cell cultures should be recorded whenever conducting toxicity tests in cell cultures with trichothecenes. Relations of chemical structures of toxins to their biological activities were investigated. With verrucarol (scirpendiol), scirpentriol, and T-2 tetraol analogues the esters generally showed significantly higher cytotoxicity. But, loss of acetyl groups at single positions did not necessarily mean decline in toxicity. With macrocyclic trichothecenes minor differences in the side chain of verrucarol esters were found to contribute to differences in biological activity. Address: Institut für Mikrobiologie, Bundesanstalt für Milchforschung, Hermann-Weigmann-Straße 1-3, D-2300 Kiel, Germany.

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Investigation of Penicillium chrysogenum Isolates for Their Suitability as Starter Cultures AA El-Banna, J Fink-Gremmels, and L Leistner Mycotoxin Research 1987, Vol. 3 (2), 77-83

One hundred twenty three isolates of P. chrysogenum were biologically tested in brine shrimp test and screened with Staphylococcus aureus for the detection of antibacterial activity. Furthermore, they were chemically examined (thin layer chromatographic method, TLC) for the synthesis of 8 mycotoxins (citrinin, cyclopiazonic acid, mycophenolic acid, patulin, penicillic acid, PR-toxin, ochratoxin A, and roquefortine). The results indicated that 85% of the tested isolates produce roquefortine and one isolate produces cyclopiazonic acid. Considering the results of the chemical assay for mycotoxins as well as the results of the brine shrimp test and the detection of antibacterial activity, 119 (97%) of the tested isolates are not suitable to be used as starter cultures for mould-fermented meats. The extracts of only 4 isolates were subjected to further biological tests in mice and the results indicated that only one isolate was non-toxinogenic. Address: Institute for Microbiology, Toxicology and Histology of the Federal Centre for Meat Research, D-8650 Kulmbach, Germany.

Cytotoxicity Evaluation of Mycotoxins by an MTT-Bioassay 1 1 2 Gerhard H Reubel , Manfred Gareis , and Werner M Amselgruber

Mycotoxin Research 1987, Vol. 3 (2), 85-96

A colorimetric tetrazolium (MTT) cleavage test was modified and established as a bioassay for the cytotoxicity of mycotoxins. Using the human erythroleukemia cell line K562 and porcine white blood cells (lymphocytes and granulocytes) we evaluated the influence of deoxynivalenol, ochratoxin A, and zearalenone on cellular MTT cleavage activity. The yellow MTT is reduced by mitochondrial enzymes of metabolically active cells into a dark blue formazan product, the optical density (OD) of which can be measured by an ELISA reader. After an exposure time of 24 hours, concentrations of deoxynivalenol and ochratoxin A as low as 0.4 μg/mL were found to inhibit significantly the cleavage activity in K562 cells. Cytotoxicity in lymphocytes and granulocytes was observed at concentrations of 0.8 up to 0.4 μg/mL for deoxynivalenol and 3.1 and 0.8 μg/mL for ochratoxin A, respectively. Zearalenone concentrations of 25.0 to 12.5 μg/mL inhibited the mitochondrial cleavage activity of lymphocytes and of K562 cells significantly, whereas in granulocytes none of the concentrations tested was proved to be toxic. Morphological findings on the ultrastructural level showed that toxin incubation (28 hours) resulted in massive cell damage. Similar alterations were observed in about 15% of control cells. This indicates, that the massive cytotoxic effect of the mycotoxin deoxynivalenol is more likely to be an unspecific than a specific one. The modified MTT cleavage assay was found to be a quick (28 hours) and efficient colorimetric test for examining the cytotoxicity of three mycotoxins. The simplicity and speed of the procedure, which allows the simultaneous testing of various parameters and the possibility of objective data analysis could establish this test as an additional bioassay for the evaluation of cytotoxicity of mycotoxins. Addresses: 1, Institute for Medical Microbiology, Infectious and Epidemic Diseases and 2, Institute of Veterinary Anatomy, Veterinary Faculty, University of Munich, Veterinärstr 13, D-8000 München 22, Germany.

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An Indirect Enzyme-Linked Immunosorbent Assay for the Detection of Diacetoxyscirpenol in Wheat and Corn Susan L Schubring and FS Chu Mycotoxin Research 1987, Vol. 3 (2), 97-106

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for quantitation of diacetoxyscirpenol (DAS) in wheat flour and corn meal. Diacetoxyscirpenol was first converted to its “b”carboxymethoxyl oxime and then conjugated to bovine serum albumin (BSA) via a water-soluble carbodiimide method. This conjugate was then coated to a microtiter plate and incubated with rabbit anti-DAS antibody and sample extract. The amount of anti-DAS antibody bound to the plate was then determined by reaction of goat anti-rabbit IgG-peroxidase complex, and subsequent reaction with substrate. Samples spiked with DAS were extracted with acetone and subjected to a simple cleanup procedure by passing them through a reversed-phase Sep-Pak C-18 cartridge. The minimum detection level for DAS was 5 picograms per assay. Average recoveries from samples of wheat flour spiked with DAS in the 1l00 ppb range were 97.5 r 17.8% for one gram samples, and 97.2 r 19.9% for fifty gram samples. Average recoveries from corn meal samples spiked in the same range were 98.8 ± 22.6% for one gram samples, and 99.7 r 17.3% for fifty gram samples. Address: Food Research Institute and Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, WI 53706, USA.

Reduction of Aflatoxin B1 Levels by Sheep Saliva Saad M Magdi Mycotoxin Research 1987, Vol. 3 (2), 107-110

This study determined the decrease of aflatoxin B1 by sheep saliva at concentrations of 150 and 300 μg aflatoxin B1/ L saliva. Analyses for aflatoxins B1, M1 and aflatoxicol (R0) were performed after 2, 4, 6, 24, and 48 hours of incubation. Aflatoxin M1 and R0 were not detected and only residues of aflatoxin B1 were found. 4 to 13% of aflatoxin B1 were decomposed by sheep’s saliva within 2 hrs and 33 to 43% of aflatoxin B1 after 24 hrs. Decomposition was affected by the aflatoxin concentration. Decrease of aflatoxin B1 at 2, 4, 6 hrs was nearly three times higher at the low concentration (150 ppb) compared to the high concentration (300 ppb). After 48 hrs incubation more than 80% of the initial aflatoxin B1 had been decomposed by the saliva. Address: Mycotoxins Lab, National Research Centre, Dokki, Cairo, Egypt.

Fusarium sporotrichioides Sherb. and Trichothecenes Associated with Fusarium-Ear Rot of Corn before Harvest 1 1 1 2 2 J Chelkowski , H Kwana , P Zajkowski , A Visconti , and A Bottalico

Mycotoxin Research 1987, Vol. 3 (2), 111-113

Fusarium sporotrichioides was found to be the predominant fungus in approximately 2 % of corn ears damaged by Fusarium species, before harvest during 1984 and 1985 in Poland. Concentrations of up to 1,714.9 mg/kg of total type-A trichothecenes (T-2 Toxin, HT-2 Toxin, Neosolaniol, T-2 Triol, and T-2 Tetraol) were found in hand-selected, heavily damaged kernels obtained from F. sporotrichioidesmolded ears. Addresses: 1, Department of Plant Pathology, Agricultural University, P-02-766 Warsaw, Poland; 2, Istituto Tossine e Micotossine da Parassiti Vegetali, Consiglio Nazionale delle Ricerche, I-70126 Bari, Italy.

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Screening for Fusarin C Production by European Isolates of Fusarium Species Ulf Thrane Mycotoxin Research 1988, Vol. 4 (1), 2-10

A total of 137 Fusarium isolates were screened for in vitro production of the mutagenic metabolite fusarin C, using a simple thin layer chromatographic method. It has been proven that Fusarium species (F. culmorum, F. graminearum, F. crookwellense, F. sporotrichioides, F. poae, F. tricinctum, and F. avenaceum) isolated from European agricultural crops and soils are able to produce fusarin C. No fusarin C production was detected among isolates of F. arthrosporioides, F. acuminatum, or F. equiseti. Results obtained by High-Performance Liquid Chromatography (HPLC) analyses of fungal extracts show that up to 26 chromatographic peaks having UV spectra similar to that of fusarin C are produced. It is not known if any of these metabolites are as mutagenic as fusarin C. Address: Department of Biotechnology, Building 221, The Technical University of Denmark, DK-2800 Lyngby, and the Biotechnical Section of the Engineering Academy of Denmark, DK-2800 Lyngby, Denmark.

Lethality of T-2 Toxin, as Applied by Various Methods, in Adult Aedes aegypti (Diptera: Culicidae) Mosquitoes Yehuda Braverman and Alan Shlosberg Mycotoxin Research 1988, Vol. 4 (1), 11-14

The lethality of T-2 toxin to adult female Aedes aegypti mosquitoes was investigated using various methods of application: Topically, by injection, and by natural feeding. The technique of topical application was found to be the most precise, whereby an LD50 of 565 ng T-2 toxin/mosquito was determined. Address: Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.

Cross-Reactivity of Antibodies Against T-2 With Deepoxide T-2 Toxin 1 2 1 Ru-dong Wei , Steven P Swanson , and FS Chu

Mycotoxin Research 1988, Vol. 4 (1), 15-19

Two types of antibodies raised against T-2 toxin, namely anti-T-2-HS-BSA and anti-3-Ac-NEOS-HSBSA, showed good cross-reactivity with deepoxy T-2 toxin. Our results indicate that the epoxide is not an important epitope for the production of antibody against T-2 toxin. Addresses: 1, Food Research Institute and Department of Food Microbiology and Toxicology, University of Wisconsin at Madison, Madison, WI 53706, USA; 2, Department of Veterinary Biosciences, University of Illinois at Urbana-Champaign, Urbana, Ill 61801, USA.

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Method for Small Routine Laboratories for the Detection of Satratoxins in Straw Samples Balázs Harrach Mycotoxin Research 1988, Vol. 4 (1), 20-24

A method is described for thin-layer chromatographic analysis of field samples for satratoxins G and H, the Stachybotrys toxins most soluble under physiological circumstances and representing the majority of Stachybotrys atra produced toxins. Besides brine shrimp bio-assay, the conversion of these macrocyclic trichothecenes to verrucarol is used to support their identification. Address: Veterinary Medical Research Institute, Hungarian Academy of Sciences, H-1581 Budapest, POB 18, Hungary.

Comparison of the Inhibition of Deoxynivalenol and T-2 toxin on Bovine and Porcine Platelet Function PA Gentry, GS Bondy, and ML Ross Mycotoxin Research 1988, Vol. 4 (1), 25-32

A platelet model system has been used to investigate the inhibitory effects of deoxynivalenol (DON, vomitoxin) and T-2 toxin, alone and in combination. In both bovine and porcine systems, the most dramatic effect observed was the instability in the platelet aggregates formed in the presence of the mycotoxins. Bovine platelets were more sensitive to the inhibitory effects of both of the mycotoxins than porcine platelets and in both species T-2 toxin was a more effective platelet inhibitor than DON. The mycotoxins may inhibit platelet function by a similar mechanism since an additive inhibitory response was observed when DON and T-2 toxin were added together to platelet suspensions. Address: Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.

Effect of Tortilla-Preparation Process on Aflatoxins B1 and B2 in Corn 1 1 2 2 HK Abbas , CJ Mirocha , R Rosiles , and M Carvajal

Mycotoxin Research 1988, Vol. 4 (1), 33-36

Naturally contaminated corn containing 450 and 54 ppb aflatoxins B1 and B2, respectively was treated with Ca(OH)2 for making tortillas. The cleaned corn and tortillas were analyzed for aflatoxins B1 and B2 by high performance liquid chromatography (HPLC) and confirmed by thin layer chromatography (TLC). The averaqe concentrations of aflatoxins B1 and B2 in the final products (tortillas) were only 40% and 28% lower than that in starting materials (corn kernels), respectively. Aflatoxins G1 and G2 were not detected in either corn or tortilla samples. Addresses: 1, Department of Plant Pathotogy, University of Minnesota, St Paul, MN 55108, USA; 2, Departamento de Botanica, lnstituto de Biologia, UNAM, Delegacion Coyoacan, 04510 Mexico, DF, Mexico.

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Synergistic Effect of Sacoglottis gabonensis Bark Extract, a Nigerian Palmwine Additive, and Ethanol on Rat Hepatic Metabolism of Aflatoxin B1 1 2 ZSC Okoye and GE Neal

Mycotoxin Research 1988, Vol. 4 (1), 37-43

Aflatoxin B1 metabolism was studied using microsomal and cytosolic fractions isolated from weanling male Fischer F344 rats given in drinking water for 7 days an aqueous extract of Sacoglottis gabonensis bark, 0.1 % ethanol solution, or a solution containing both extract and ethanol ad libitum. Microsomal production of aflatoxin B1-dihydrodiol, aflatoxin Q1, aflatoxin M1, aflatoxin P1 and protein-aflatoxin adduct formation, and cytosolic aflatoxin B1-glutathion conjugation were assayed. Pretreatment with the extract alone or together with ethanol caused significant increases in aflatoxin M1 production as compared to controls given only water, but aflatoxin Q1 production was enhanced only by pretreatment with both extract and ethanol. All the three treatments caused significant reductions in liver glutathione content. The highest aflatoxin B1 metabolising activity as determined by aflatoxin M1 and aflatoxin Q1 production was observed in rat pretreated with both ethanol and the extract, suggesting synergism. The findings suggest that at relatively mild doses, S. gabonensis extract alone or in concert with ethanol may influence response to aflatoxin. Addresses: 1, Department of Biochemistry, University of Jos, Jos, Nigeria; 2, MRC Toxicology Unit, Medical Research Council Laboratories, Carshalton, Surrey, United Kingdom.

Detection and Quantification of Patulin and Griseofulvin by High Pressure Liquid Chromatography in Different Strains of Penicillium Griseofulvum Dierckx M Jiménez1, V Sanchis2, R Mateo3, and E Hernández1 Mycotoxin Research 1988, Vol. 4 (2), 59-66

Patulin and griseofulvin production by twelve strains of Penicillium griseofulvum Dierckx, eleven of which were isolated from pistachio (Pistacia vera) nuts and the other was supplied by the Spanish Collection of Type Culture, was investigated. Six strains of the eleven isolated had ability to produce patulin and griseofulvin in Yes medium. All the strains studied had no ability to produce patulin in Wickerham medium. Griseofulvin production was significant in both media but higher in Wickerham. These metabolites were separated and determined in the chloroform extracts of cultures by high performance liquid chromatography with ultraviolet detection. The best conditions were: acetonitrile water (45:55) as mobile phase, a flow rate of 2.0 mL/min and a μBondapack C18 column. Addresses: 1, Department of Biotechnology, Polytechnic University of Valencia, Camino de Vera, E-46022 Valencia, Spain; 2, Department of Microbiology, ETSIA, Polytechnic University of Catalunya, Lleida, Spain; 3, Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia, Dr Moliner 50, Burjassot, Valencia, Spain.

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Production of Fusarenon-X, Nivalenol, and Zearalenone by Gibberella zeae Isolates, and Their Toxicity in Fibroblasts and Rats HK Abbas and CJ Mirocha Mycotoxin Research 1988, Vol. 4 (2), 67-74

Three isolates of Gibberella zeae, the perfect stage of Fusarium graminearum, were isolated from ground corn cultures obtained from Taiwan in 1985 and identified as Gibberella zeae I-1, G. zeae I5, and G. zeae I-7. The isolates were grown on a solid rice medium and extracts prepared with 75 % aqueous methanol. The extracts were examined for toxicity in the following systems: (1) cytotoxicity to cultured normal human diploid skin fibroblasts and mouse fibroblasts; and (2) toxicity to rats of unextracted cultures. The three extracts were highly cytotoxic as indicated by the ability to cause death and disintegration of 3T3 Swiss mouse fibroblasts and human diploid skin fibroblasts during 3 to 4 days in culture. The unextracted cultures of the isolates were highly toxic to rats, causing hemorrhage of tissues (bladder, stomach, and intestine), uterine enlargement, small thymuses, small spleens, weight loss, and death. The extracts were tested for production of trichothecenes (nivalenol and fusarenon-X) and zearalenone on rice grains. Production of the three mycotoxins was greater at room temperature than in the cold room. Detection of the three mycotoxins from the cultures was variable, ranging from 273 to 8l7 ppm for nivalenol, 268 to 662 ppm for fusarenon-X, and 162 to 1095 ppm for zearalenone at room temperature, and 159 to 413 ppm for nivalenol, 113 to 125 ppm for fusarenon-X and 44 to 202 ppm for zearalenone in the cold room (10°C). Address: Department of Plant Pathology, University of Minnesota, St Paul, Minnesota 55108, USA.

A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay of Aflatoxin B1 in Peanut Products O Kawamura1, S Nagayama1, S Sato1, K Ohtani1, I Ueno2, and Y Ueno1 Mycotoxin Research 1988, Vol. 4 (2), 75-88

An improved enzyme-linked immunosorbent assay (ELISA) combined with monoclonal antibody (MAb) and one-step extraction method was established for the estimation of aflatoxin B1 (AFB1) in a peanut product. AFB1 was converted to AFB1-oxime, and then conjugated with bovine serum albumin (BSA). Spleen cells from mice immunized with AFB1-BSA conjugates were fused with myeloma cells. After double selection with AFB1-ovalbumin (OVA) and carbodiimide-modified OVA, five stable hybridoma cells secreting anti-AFB1 MAbs (AF 1, AF 2, AF 3, AF 4, and AF 5) were cloned. Using these anti-AFB1 MAbs, we developed the indirect competitive ELISA (cELISA) with alkaline phosphatase (ALP) - labeled sheep anti - mouse IgG as marker and the direct cELISA with AFB1oxime horse-radish peroxidase (POD) as marker. The minimum detectable limits of the indirect cELISA with AF 1, 2, 3, 4, and 5 were 5, 5, 5, 5, and 50 pg of standard AFB1 per assay, respectively, and those of the direct cELISA with AF 1, 3, 4, and 5 were 2.5, 5, 25, and l00 pg of standard AFB1 per assay, respectively. The cross reactivity of each toxins with these MAbs in the indirect cELISA was as follows: (a) AF 1 and AF 2 were reactive with AFB2 as well as AFB1, weakly with AFG2 > AFG1 > aflatoxicol II (COL II) > aflatoxicol I (COL 1) and less weakly with other aflatoxins; (b) AF 3 and AF 4 were reactive with COL II as well as AFB1, weakly with COL I > AFQ1 and less weakly with others; (c) AF 5 was AFQ1 as well as AFB1, weakly with COL II > AFG2 > COL I and less weakly with others. The 60% aqueous methanol extracts of oil-roasted blanched peanuts (“butter peanut“), naturally contaminated with AFB1, were assayed by the direct cELISA without further purification. The direct cELISA with the most sensitive MAb AF 1 was allowed to determine 1 ng of AFB1 per g samples. Addresses: 1, Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Tokyo 162, Japan; 2, The Institute of Medical Sciences, University of Tokyo, Shiroganedai, Tokyo 108, Japan.

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Levels of Aflatoxin B1, Bacteria and Fungi in Feed and Food in 1971 - 1987 2

2

2

2

EL Strzelecki, U Gsiorowska , M Gorazdowska , B Cader-Strzelecka , and M Paweczak Mycotoxin Research 1988, Vol. 4 (2), 89-96

Totally 39 % out of 8371 feed and their component samples were contaminated by aflatoxin B1. Mean contamination was 36 μg/kg with maximum yield 10100 μg/kg. Contamination of samples by total count of organisms, mean contamination and maximum yield, respectively was: 1) bacteria 99 %, 2.2x106, 2.4x108; 2) proteolytic bacteria 94 %, 1.2x105, 3.0x106; 3) moulds 98 %, 1.3x105, 9.0x106; 4) yeasts 44 %, 3.3x104, 3.6x106. The samples were contaminated in 92 % by Aspergillus spp., in 71 % by Aspergillus flavus, in 83 % by Penicillium spp., and in 20 % by Fusarium spp. with mean contamination 8.3x104, 1.1x103, 4.2x104, 5.0x103‚ and maximum yield 6.8x106, 1.0x105, 5.0x106, 1.5x106, respectively. Totally 8.5 % of strains were aflatoxinogenic and 4.4 % of the strains were isolated from feed and 21% of the strains from grain/nut. Address: Mycology and Mycotoxicology Laboratory, Veterinary Hygiene Research Station, 249 Kartuska St, PL-80125 Gdask, Poland.

Deoxynivalenol and 3-acetyldeoxynivalenol and Fusarium Species in Winter Triticale J Perkowski1, J Chelkowski2, and W Wakuliski2 Mycotoxin Research 1988, Vol. 4 (2), 97-100

Fusarium species infecting heads of Triticale and mycotoxins presence in infected kernels and chaff were studied during two seasons. The most important species observed on infected heads were in 1986 F. avenaceum (39 %), F. nivale (21 %), F. culmorum (20 %), F. graminearum (14 %), and others (6 %). In 1987 after long and snowy winter F. nivale dominated (64 %), followed by F. avenaceum (24 %), F. culmorum (6 %), and F. graminearum (5 %). The mycotoxins deoxynivalenol (DON) and 3-acetyl DON were present in all 11 subsamples of kernels from heads infected by F. culmorum and/ or F. graminearum (1.6-16.4 mg and 0.7-2.4 mg/kg, respectively). Chaff from the same subsamples contained 9.9-33.2 mg/kg of DON and 5.2-16.0 mg/kg of 3-AcDON. Kernels with visible Fusariumdamage contained 2.4-31.2 mg/kg of DON and 1.2-6.0 mg/kg of 3-AcDON. Remaining part of kernels without symptoms of visible Fusarium-damage contained only DON in an amount of 0.9-5.9 mg/kg. Addresses: 1, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University, 02-766 Warsaw, Poland.

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Toxicity of Emestrin, a New Macrocyclic Dithiodioxopiperazine Mycotoxin, to Mitochondrial Function 1 1 2 2 2 3 Kawai K , Ishizaki K , Nakamaru T , Hisada K , Nozawa Y , Kawai KI

Mycotoxin Research 1989, Vol. 5 (1), 2-8

The effect of emestrin, a new macrocyclic epidithiodioxopiperazine mycotoxin from several Emericella species, on mitochondrial reactions was studied using isolated rat liver mitochondria to gain insight into the molecular mechanism for its in vivo toxicity to rat and mouse. Emestrin was found to inhibit ATP synthesis in mitochondria causing an uncoupling of oxidative phosphorylation and a depression of respiration in isolated mitochondria. In addition to these effects on mitochondrial respiration, emestrin elicited a drastic structural alteration (swelling) of mitochondria as observed in the in vivo system. The mitochondrial swelling was significantly enhanced by the subsequent addition of calcium ion. Emestirin B, in which dithio group is replaced by trithio group, exerted an uncoupling effect on oxidative phosphorylation without accompanying such depressive effect on state 3 respiration as observed for emestrin. Addresses: 1, Faculty of Home Economics, Chukyo Women‘s University, Ohbu 474, Aichi, Japan; 2, Department of Biochemistry, Gifu University School of Medicine, Gifu 500, Japan; 3, Faculty of Pharmaceutical Science, Hoshi University, Tokyo 142, Japan.

Incidence of Aflatoxin, Zearalenone, and Deoxynivalenol on Corn in Argentina 1 2 2 Chulze S , Bertinetti C, Dalcero A, Etcheverry M, Farnochi C, Torres A, Rizzo I , Varsavsky E

Mycotoxin Research 1989, Vol. 5 (1), 9-12

The study of the incidence of aflatoxins, zearalenone, and deoxynivalenol was the aim of this work. This investigation was carried out upon recently harvested corn samples collected in the Departamento of Río Cuarto, Province of Córdoba, Argentina. 150 samples of corn were analized to investigate aflatoxins B1, G1, and zearalenone contamination. Out of these 150 samples 58 were selected and deoxynivalenol was examined. The incidence value for aflatoxin B1 was of 3.3% (5 samples) aflatoxin G1 1.3% (2 samples), and zearalenone 6% (9 samples). The analysis of the 58 samples showed that 24% of them were contaminated with deoxynivalenol. Addresses: 1, Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico Químicas y Naturales, Universidad Nacional de Río Cuarto, Estafeta Postal No 9-5800 Río CuartoCórdoba, Argentina; 2, Instituto Nacional de Farmacología y Bromatología (INFYB), Buenos Aires, Argentina.

Production of Neosolaniol Monoacetate by an Undescribed Fusarium Species Resembling F. camptoceras 1 1 1 2 Sydenham EW , Thiel PG , Marasas WFO , and Lamprecht SC

Mycotoxin Research 1989, Vol. 5 (1), 13-19

Cultures of 12 South African isolates of an undescribed Fusarium species resembling but distinct from F. camptoceras were analysed for the presence of diacetoxyscirpenol (DAS), neosolaniol monoacetate (NMA), and T-2 toxin by capillary gas chromatography utilizing electron capture detection. No DAS or T-2 toxin could be detected in any of the cultures of the isolates. NMA was, however, detected in 10 of the 12 isolates at levels ranging from 310 to 2060 ng/g. The method used, was primarily developed for the determination of DAS and T-2 toxin in fungal cultures and grain samples but was found to be suitable for the coextraction of NMA at an average recovery of 80.8%, with a detection limit in the order of 100 ng/g. Supportive evidence for the presence of the NMA was obtained by capillary gas chromatography / mass spectrometry. Regarded as a relatively rare trichothecene, NMA has never 191

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1989

been reported to occur naturally and has previously been shown to be produced by only a few Fusarium strains. Addresses: 1, Research Institute for Nutritional Diseases, South African Medical Research Council, PO Box 70, Tygerberg 7505, South Africa; 2, Plant Protection Research Institute, Private Bag X5017, Stellenbosch 7600, South Africa.

Functional Alterations Induced by Fusarium T-2 Toxin in Madin-Darby Bovine Kidney (MDBK) and Primary Fetal Bovine Kidney (PFBK) Cultures Masao Yoneyama and Raghubir P Sharma Mycotoxin Research 1989, Vol. 5 (1), 21-29

We recently reported that primary fetal bovine Kidney (PFBK) cells were consistently more sensitive to the cytotoxic effects of Fusarium T-2 toxin than Madin-Darby bovine kidney (MDBK) cells in culture. The present report examined the influence of T-2 on selected biochemical parameters of these two culture types. T-2 toxin inhibited incorporation of labeled thymidine, uridine, and leucine in both culture types; at lower concentrations of the toxin, PFBK cells were affected to a greater extent than MDBK cells. T-2 toxin inhibited both the transport of thymidine as well as thymidine incorporation into macromolecules in MDBK cells during initial periods, but did not affect uridine incorporation. The cellular enzymes, K+- dependent phosphatase and succinic dehydrogenase were inhibited in MDBK but not in PFBK cultures; acid phosphatase was not influenced in either culture types. In a cell-free system none of the above enzymes were affected by T-2 until the toxin concentration exceeded 10-5M. Address: Toxicology Program, Utah State University, Logan, UT 84322-5600, USA.

Isolation of 15-Acetoxyscirpendiol from Culture of Fusarium poae on Corn Evidente A1, Randazzo G2, Visconti A3, and Bottalico A3 Mycotoxin Research 1989, Vol. 5 (1), 30-34

The main toxic metabolites of a strain of Fusarium poae, isolated from oats, were diacetoxyscirpenol (DAS) and a monoacetoxyscirpendiol. An accurate 1H and 13C NMR analysis allowed to identify the monoacetoxyscirpendiol as the 15-acetoxy-3,4-dihydroxy -12,13-epoxytrichothec-9-ene (15acetoxyscirpendiol). This is the first report on the production of 15-acetoxyscirpendiol by Fusarium poae. Addresses: 1; Istituto di Chimica, Università della Basilicata,Via N Sauro, 85, I-85100 Potenza, Italy; 2, Istituto di Chimica Agraria, Università di Napoli, Via Università 100, I-80055 Portici (NA), Italy; 3, Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Via G Amendola 197/F, I-70126 Bari, Italy.

A Microbiological Assay for Sterigmatocystin, T-2 Toxin and Zearalenone 1 2 1 2 Tawfek NF , El-Sayed AM Abd-Alla , Sharaf OM , and Badawey A

Mycotoxin Research 1989, Vol. 5 (1), 35-40

A survey was done to find microorganisms useful for assaying sterigmatocystin; T-2 toxin and zearalenone. Staphylococccus aureus was found to be sensitive to T-2 toxin and zearalenone; Bacillus cereus was found to be sensitive to T-2 toxin only; and Escherichia coli was sensitive to sterigmatocystin. The reponse of the organisms to sterigmatocystin; T-2 toxin and zearalenone was found to be linear between 4 and l00 μg with sterigmatocystin to E.coli; between 2 and 25 μg with T-2 toxin to 192

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Staph. aureus and B. cereus; and between 4 and l00 μg with zearalenone to Staph. aureus. The lower limits of sensitivity of the test were 2 μg T-2 toxin and zearalenone, and 4 μg sterigmatocystin. The assay is rapid (15-17 hrs); simple and inexpensive; and can be used to verify the toxicity of samples and to confirm thin layer chromatographic results. Address: 1, Food Technology & Dairying Lab and 2, Mycotoxins Lab, National Research Centre, Dokki, Cairo, Egypt.

lmproved Methodology for Detecting Aflatoxin Production Quantitatively in Natural Media MA Moreno, A Olivares, and G Suárez Mycotoxin Research 1989, Vol. 5 (2), 51-56

This report describes a simple, rapid and quantitative method for screening the aflatoxin production by moulds of the Aspergillus flavus group, using a natural media (moist wheat or rice), and a single chloroform extraction for aflatoxins. The aflatoxins were detected and quantified by thin layer chromatography. The methodology proposed was useful in detecting aflatoxin production on the 3rd day of incubation in more than 85% of the strains studied (both culture collection strains and field isolates). Address: UD Microbiología e Inmunología, Departamento Patología Animal I (Sanidad Animal), Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.

Aflatoxin in Human and Camel MiIk in Abu Dhabi, United Arab Emirates AM Saad1, AM Abdelgadir1, and MO Moss2 Mycotoxin Research 1989, Vol. 5 (2), 57-60

During an initial survey, using thin layer chromatography, 10 of 64 samples of mothers‘ breast milk, collected from donors at the Corniche Maternity Hospital and the Al-Nehyan Clinic for Maternity and Childhood, were found to contain aflatoxin M1 at concentrations ranging from 0.3 to 1.3 ng mL-1. A second survey using HPLC showed aflatoxin M1 at concentrations ranging from 7 to 23 pg mL-1 in all of the 15 samples collected. 6 of 20 samples of camel milk collected from several sources in Abu Dhabi were also found to contain aflatoxin M1 at levels ranging from 0.25 to 0.8 ng mL-1. Addresses: 1, Food Control Laboratory, PO Box 3902, Abu Dhabi, United Arab Emirates; 2, Department of Microbiology, University of Surrey, Guildford, Surrey, GU2 5XH, United Kingdom.

The Yields of Diacetoxyscirpenol Produced by Fusarium sambucinum Cultures Isolated from Potato Tubers and Their Toxicity to Brine Shrimps (Artemia salina) 1 1 2 J Perkowski , E Foremska , and D Latus-Zitkiewicz

Mycotoxin Research 1989, Vol. 5 (2), 61-67

The amount of diacetoxyscirpenol (DAS) produced by 14 isolates of Fusarium sambucinum under laboratory conditions has been examined. The isolates were obtained from potato tubers with dry rot symptoms. Yields of DAS ranged from 20 to 330 ng/kg of wheat grain culture (chemical analysis  TLC). Chemical analysis and biological tests showed very similar results. Addresses: 1, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland; 2, Institute of Food Technology of Plant Origin, Agricultural University, 60-624 Pozna, Poland.

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Isolation and Structure Elucidation of Isoaltenuene, a New Metabolite of Alternaria alternata 1 1 1 2 Visconti A , Bottalico A , Solfrizzo M , and Palmisano F

Mycotoxin Research 1989, Vol. 5 (2), 69-76

Isoaltenuene, a previously unknown Alternaria metabolite has been isolated from a rice culture of Alternaria alternata and purified by semipreparative HPLC. The assigned structure, elucidated by UV, IR, MS, and NMR spectroscopy, was 2´,3´,4´,5´-tetrahydro-3,4´,5´-trihydroxy-5-methoxy-2´methyldibenzo()-pyrone and corresponded to a diasteroisomer of altenuene with inverted configuration at C-2´. Isoaltenuene showed a minor phytotoxic activity on tomato leaves at levels of 20 μg/spot and no antifungal activity on Geotrichum candidum (up to 20 μg/disk). Addresses: 1, Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Via Amendola 197/F, I-70126 Bari, Italy; 2, Dipartimento di Chimica, Università, 4 Trav. 200 Re David, I-70126 Bari, Italy.

Natural Occurrence of Alternaria Mycotoxins in the Grain and Chaff of Cereals 1 2 2 J Grabarkiewicz-Szczsna , J Chelkowski , and P Zajkowski

Mycotoxin Research 1989, Vol. 5 (2), 77-80

Samples of wheat and rye heads with evident black discoloration were collected in 1986 and 1987. Four fungal genera colonized such heads: Alternaria, Cladosporium, Drechslera, and Epicoccum. In chaff of wheat (19 %) and rye (10 %) samples alternariol was present in amounts up to 1.8 mg/kg, and alternariol methyl ether up to 0.51 mg/kg. In 1 out of 21 wheat samples alternariol was also present in kernels (0.59 mg/kg). Addresses: 1, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University, 02 -766 Warsaw, Poland.

Mould and Mycotoxin Contamination of Stored Corn in Turkey G Özay and D Heperkan Mycotoxin Research 1989, Vol. 5 (2), 81-89

A total of 167 corn samples, including imported and locally grown corn, were obtained from various regions and store houses in Turkey and surveyed for mould occurrence and mycotoxin content. The mould contamination level was 105106 colonies/g. A. flavus, A. niger, Fusarium oxysporum, Penicillium variable, and Rhizopus spp. However, the dominant flora showed significant differences between the imported and domestic corns. Aflatoxin B1 was found in 16% of the samples ranging from 274 μg/kg. Ochratoxin A and sterigmatocystin were found at minimum detection levels. Mycotoxin production characteristics of mould isolates were also determined. Address: TUBITAK, Marmara Scientific and Industrial Research Institute, Food Technology and Nutrition Dept, PO Box 21, 41401 Gebze, Kocaeli, Turkey.

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Determination of Aflatoxin Levels in Peanut Butter using HPLC and ELISA Procedures: Inter-Laboratory Comparison AL Patey, M Sharman, and J Gilbert Mycotoxin Research 1990, Vol. 6 (1), 2-6

Six laboratories analyzed portions of the same aqueous acetonitrile extracts of three peanut butters for aflatoxin concentrations by an HPLC procedure (using immunoaffinity column clean-up) and by an ELISA procedure. The extracts were from a nominal “blank“ peanut butter, a peanut butter naturally contaminated with aflatoxins (mostly B1) and from a “blank“ peanut butter to which equal amounts of aflatoxin B1, B2, G1 and G2 standards had been added. Similar results for the HPLC and ELISA procedures were obtained for the blank (means 2.8 and 4.0 μg/kg, respectively) and naturally contaminated (means 26.0 and 25.9 μg/kg, respectively) peanut butters. However, the result by ELISA (mean 16.7 μg/kg) for the spiked peanut butter was much lower than that obtained by HPLC (mean 28.3 μg/kg). Address: Ministry of Agriculture, Fisheries and Food, Food Science Laboratory, Colney Lane, Norwich, NR4 7UQ, United Kingdom.

Natural Occurrence of Deoxynivalenol, 3-Acetyl-Deoxynivalenol, 15-AcetylDeoxynivalenol, Nivalenol, 4,7-Dideoxynivalenol, and Zearalenone in Polish Wheat J Perkowski1, RD Plattner2, P Goliski1, RF Vesonder2, and J Chelkowski3 Mycotoxin Research 1990, Vol. 6 (1), 7-12

Three wheat samples collected in 1987 in Central Poland and naturally infected with Fusarium spp. were analyzed for the presence of Fusarium spp and Fusarium toxins. Heads were separated into three fractions: kernels with visible Fusarium damage, healthy looking kernels, and chaff + rachis. The samples contained deoxynivalenol (2.040.0 μg/g), nivalenol (0.01 μg/g), 4,7-dideoxynivalenol (0.100.15 μg/g), 15-acetyldeoxynivalenol (0.102.00 μg/g), 3-acetyldeoxynivalenol (0.10 μg/g), and zearalenone (0.012.00 μg/g). This is the first report about 15-acetyldeoxynivalenol in European wheat and the co-occurrence of 3-acetyldeoxynivalenol and 15-acetyldeoxynivalenol in the same sample of contaminated cereals. Addresses: 1, Department of Chemistry, Agricultural University of Pozna, ul Wojska Polskiego 75, 60-625 Pozna, Poland; 2, Northern Regional Research Center, Agricultural Research Service, US Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604, USA; 3, Department of Plant Pathology, Agricultural University of Warsaw, ul Nowoursynowska 166, 02-766 Warsaw, Poland.

Evaluation of Trichothecene and Nontrichothecene Mycotoxins Produced by Fusarium in Soybeans HK Abbas and U Bosch Mycotoxin Research 1990, Vol. 6 (1), 13-20

Each of 12 cultures of Fusarium, comprising four species, isolated from moldy soybeans suspected of being involved in illness of wild geese, were grown separately in autoclaved moist rice, in autoclaved moist soybeans, and in surface sterilized - disinfected soybeans, assayed for various mycotoxins, and fed to rats. Four additional cultures that produced known toxins on rice were also grown on soybeans as controls. All isolates, except one of F. moniliforme, grown in rice resulted in weight loss of rats, and that one resulted in weight gain; 12 of the isolates caused death. One isolate of F. poae grown in soybeans caused death when consumed by rats, but none of the other 15 resulted in weight loss or overt injury. Much larger amounts of zearalenone, deoxynivalenol (DON), T-2 toxin, neosolaniol, T-2 tetraol, wortmannin, and moniliformin were produced by the cultures on rice than on soybeans, but 195

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more HT- 2 toxin was produced by one isolate of F. poae grown on soybeans than when grown on rice. Soybeans appear to be a poor substrate for elaboration of most of the toxins produced by the isolates tested. Address: Department of Plant Pathology, University of Minnesota, St Paul, MN 55108, USA.

Isolation, Identification and Toxicological Characterization of TSS 1, a New Mycotoxin of the Rosenane Class M Schäfer1, J Hengstler2, S Löffler2 and P Flesch3 Mycotoxin Research 1990, Vol. 6 (1), 21-30

11--hydroxy-7-deoxy-rosenonolactone (TSS 1), a product of the pathogenic fungus Trichothecium roseum (Moniliaceae) was isolated from culture medium extracts and completely described in its structure by spectroscopical methods. TSS 1 was classified as a representative of the lactone series of the rosenane class and as a structural isomer to Rosenololactone and Rosololactone. TSS 1 showed toxic effects in the growth inhibition test to Escherichia coli (EC 50: 10 μg/mL) and Bacillus subtilis (EC 50: 17 μg/mL), inhibited fermentation of yeast (EC 50: 2.8 μg/mL) and suppressed motility of Artemia salina larvae (EC 50: 45 μg/mL). Rosenonolactone, the best known representative of that mycotoxin class, showed only a very weak biological activity (EC 50: 200 μg/mL), despite the high degree of structural similarity compared to TSS 1. This investigation demonstrates that a small difference in the chemical structure between two metabolites of the same organism can strongly enhance the toxicity of a substance. Addresses: 1, Firma E Merck, D-6100 Darmstadt, Germany; 2, Institut für Toxikologie, Universität Mainz, D-6500 Mainz, Germany; 3, Institut für Biochemie, Universität Mainz, D-6500 Mainz, Germany.

The Fate of the Fusarium Mycotoxin Zearalenone in Maize Cell Suspension Cultures G Zill, G Engelhardt, B Wohner, PR Wallnöfer Mycotoxin Research 1990, Vol. 6 (1), 31-40

The Fusarium mycotoxin zearalenone was transformed in cell suspension cultures of Zea mays giving - and -zearalenol and the -D-glucosides of zearalenone and - and -zearalenol. The structure of zearalenone-4--D-glucopyranoside was determined by liquid chromatography-mass spectrometry and specific hydrolysis with -glucosidase. - and -zearalenol and their glucosides were identified by cochromatography using TLC and HPLC and glucosidase-treatment. Up to 50 % of the mycotoxin added was bound to a non extractable or “bound“ residue fraction. After treating this residue by a sequential cell wall fractionation procedure, zearalenone was found to be bound mainly to starch, hemicellulose, and lignin fractions. Address: Bayerische Landesanstalt für Ernährung, Menzingerstraße 54, D-8000 München 19, Germany.

Moniliformin Production by Fusarium Species J Chelkowski1, M Zawadzki1, P Zajkowski1, A Logrieco2, and A Bottalico2 Mycotoxin Research 1990, Vol. 6 (1), 41-45

A total of 132 Fusarium isolates belonging to 19 species sensu Nelson et al. (1983) originating from Poland, Italy, and international cultures collections were examined for their ability to produce mycotoxin moniliformin. Moniliformin was produced by the following isolates: F. acuminatum Ell & Ev: 2 196

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1990

out of 2, 1302670 mg/kg; F. avenaceum (Fr) Sacc: l8 out of 18, 702670 mg/kg; F. anthophilum (A Braun) Wollenw: 1 out of 3, 200 mg/kg; F. dlamini Marasas et al: 2 out of 3, 130470 mg/kg; F. oxysporum Schlecht emend Snyd Hans: 4 out of 9, 130270 mg/kg; F. proliferatum (Matsushima) Nirenberg: 3 out of 7, 130400 mg/kg; F. solani (Mart) Appel & Wollenw: 1 out of 14, 670 mg/kg; F. subglutinans (Wollenw & Reinking) Nelson et al: 8 out of 20, 701660 mg/kg; F. tricinctum (Corda) Sacc: 2 out of 9,1301330 mg/kg. In cultures of F. beomiforme Nelson, Toussoun & Burgess, F. chlamydosporum Wollenw & Reinking, F. compactum / Wollenw / Gordon, F. equiseti / Corda / Sacc, F. poae / Peck / Wollenw, F. moniliforme Sheldon, F. napiforme Marasas, Nelson & Rabie, F. nygamai Burgess & Timbold, F. polyphialidicum Marasas et al, F. sporotrichioides Sherb moniliformin was not detected. The highest amounts of moniliformin by F. avenaceum using solid substrate were formed on rice and lower on oats kernels. Addresses: 1, Department of Plant Pathology, Agricultural University, 02766 Warsaw, Poland; 2, Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, 70126 Bari, Italy.

Production of DON-Related Trichothecenes by Fusarium graminearum Schw from Krasnodarski Krai of the USSR AN Leonov, GP Kononenko, and NA Soboleva Mycotoxin Research 1990, Vol. 6 (2), 54-60

34 Fusarium graminearum Schw isolates produced 4-deoxynivalenol to form significant amounts of 4,7-dideoxynivalenol and lesser amounts of 4-deoxynivalenol monoacetates on grain substrates in vitro. This is the first report on the capability a large group of naturally occurring isolates to produce 4,7-dideoxynivalenol. The average levels of 4,7-dideoxynivalenol on rice, corn, barley, and wheat as a substrate were respectively 26.8, 14.0, 12.8, and 10.5 % of the level of 4-deoxynivalenol. 4,7-dideoxynivalenol was present in all examined naturally contaminated wheat kernel samples at levels of 1.7 to 7.9 % of the level of 4-deoxynivalenol. These findings suggest that more attention should be given to the occurrence of 4,7-dideoxynivalenol in cereals. Address: Institute of Veterinary Sanitation, 123022 Moscow, Zvenigorodskoe shosse, 5, USSR.

Comparative Effects of Trichothecene Mycotoxins on Bovine Platelet Function: Acetyl T-2 toxin, a More Potent Inhibitor than T-2 Toxin KM Grandoni1, PA Gentry1, BJ Holub2, and B Yagen3 Mycotoxin Research 1990, Vol. 6 (2), 61-66

The effects of the trichothecene mycotoxins (acetyl T-2 toxin, T-2 toxin, HT-2 toxin, palmityl T-2 toxin, diacetoxyscirpenol (DAS), deoxynivalenol (DON), and T-2 tetraol on bovine platelet function were examined in homologous plasma stimulated with platelet activating factor (PAF). The mycotoxins inhibited platelet function with the following order of potency: acetyl T-2 toxin > palmityl T-2 toxin = DAS > HT-2 toxin = T-2 toxin. While T-2 tetraol was completely ineffective as an inhibitor, DON exhibited minimal inhibitory activity at concentrations above 10 x 10-4 M. The stability of the platelet aggregates formed was significantly reduced in all mycotoxin treated platelets compared to that of the untreated PAF controls. It is suggested that the increased sensitivity of PAF stimulated bovine platelets to the more lipophilic mycotoxins may be related to their more efficient partitioning into the platelet membrane compared to the more hydrophilic compounds. Addresses: 1, Department of Biomedical Sciences, and 2, Department of Nutritional Sciences, College of Biological Sciences, Ontario Veterinary College, University of Guelph, Guelph, Ontario, N1G 2W1, Canada; 3, Department of Natural Products, School of Pharmacy, The Hebrew University of Jerusalem, 91120, Israel.

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Bioconversion of Aflatoxin G1 to Aflatoxin B3 by Selected Fungi 1

2

N Lisker , GS Zhang , and Fun S Chu

2

Mycotoxin Research 1990, Vol. 6 (2), 67-72

Aflatoxin G1 (AFG1) was transformed into aflatoxin B3 (AFB3) by the fungi Rhodotorula sp., Sporobolomyces sp., Rhizopus oryzae NRRL 395, Pythium ultimum, Aspergillus terreus, A. clavatus and Penicillium frequentans grown in a medium containing AFG1, Difco potato dextrose broth, yeast extract, and peptone both in liquid shaken cultures and in solid static cultures at 25 °C in the dark. A maximum rate of transformation of 10 % was obtained after 2 to 3 weeks of incubation. The transformation was correlated with an increase in the pH of the media from 5.75.9 to 8.38.8. Saccharomyces cerevisiae also transformed AFG1 into AFB3, but at a slower rate; the pH of the media did not reach above 8.0 until 5 weeks after incubation. No transformation was observed when A. niger and P. chrysogenum were tested; in both cases, no increase in pH was noticed. However, some transformation of AFG1 to AFB3 by both fungi was observed when the initial pH of the media was adjusted to 9.0. The rate of transformation increased to 1520 % in the static culture where the same medium was adsorbed onto vermiculite and Rhizopus and Aspergilli gave the highest increase in AFB3 yield. Addresses: 1, Department of Agronomy and Natural Resources, Agricultural Research Organization, The Volcani Center, Bet Dagan 50250, Israel; 2, Food Research Institute and Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, WI 53706, USA. Production and Characterization of Polyclonal and Monoclonal Antibodies against Aflatoxin B1 Oxime-BSA in an Enzyme-Linked Immunosorbent Assay CM Ward1, AP Wilkinson1, S Bramham1, HA Lee1, HWS Chan1, GW Butcher2, A Hutchings2, 1 and MRA Morgan Mycotoxin Research 1990, Vol. 6 (2), 73-83

From a single aflatoxin B1 oxime-bovine serum albumin conjugate, polyclonal and monoclonal antibody preparations were produced. The four rabbit polyclonal antisera were specific for aflatoxin B1 in a microtitration plate enzyme-linked immunosorbent assay. The monoclonal antibodies showed a wide range of differing specificities, recognizing, for example, aflatoxins B1, B2, G1, and G2; B1 and B2; B1 and G1; and G2 alone. No antibody preparations reacted with aflatoxin M1. The significance of these results to the strategy of anti-aflatoxin antibody production for use in quantitative enzyme-immunoassays is discussed. Addresses: 1, AFRC Institute of Food Research, Colney Lane, Norwich, NR4 7UA, United Kingdom; 2, Monoclonal Antibody Centre, Department of Immunology, AFRC Institute of Animal Physiology and Genetics Research, Cambridge Research Station, Babraham, Cambridge CB2 4AT, United Kingdom. Fumonisin B1: Isolation from Corn Culture, and Purification by High Performance Liquid Chromatography R Vesonder, R Peterson, R Plattner, and D Weisleder Mycotoxin Research 1990, Vol. 6 (2), 85-88

A method is described to isolate fumonisin B1 (FB1) from corn cultured for 18 days at 25 °C with Fusarium moniliforme. Cultured corn was extracted with aqueous methanol and purified with XAD-2 column chromatography and high performance liquid chromatography (HPLC). About 450 mg of FB1 were obtained from 800 g cultured corn. Its identity was established by fast-atom bombardment (FAB) mass spectrometry, and infrared spectrum and nuclear magnetic spectrum. Its purity was estimated to be 95% by gas chromatography/mass spectrometry (GC/MS). Address: Mycotoxin Research, US Department of Agriculture, Agricultural Research Service, Northern Regional Research Center, 1815 North University Street, Peoria, IL 61604 USA. 198

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1990

Deoxynivalenol and Nivalenol in Wheat and By-Products in Argentina A Torres, S Chulze, A Dalcero, M Etcheverry, and C Farnochi Mycotoxin Research 1990, Vol. 6 (2), 89-92

The deoxynivalenol and nivalenol contamination in wheat and by-products obtained through milling was analyzed by Trucksess method slightly modified in the proportion of acetonitrile-water (3:1). Only one sample of wheat showed deoxynivalenol contamination, 1,200 μg/kg. No samples obtained in different stages of the milling were contaminated with deoxynivalenol or nivalenol. In the comercial wheat flours the levels found ranged between 400 and 800 μg/kg, as follows: 400 μg/kg, 5 samples; 800 μg/kg, 1 sample. Address: Departamento de Microbiología, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina.

Formation of Ascochitine by Plant Pathogens of the Genus Ascochyta 1 2 2 E Foremska , J Marcinkowska , and J Chelkowski

Mycotoxin Research 1990, Vol. 6 (2), 93-97

Strains of fungi isolated from pulse crops: pea (Pisum sativum) and faba beans (Vicia faba) plants with symptoms of Ascochyta blight, footrot and stems lesions have been examined under laboratory conditions for their ability to produce ascochitine and metabolites toxic to Artemia salina. Both Ascochyta pisi Lib and Ascochyta fabae LK Jones isolates formed ascochitine in yields of 20480 mg/kg. The highest yield of ascochitine was produced on rice and the lowest on maize grain. Ascochyta pinodes and Phoma medicaginis var. pinodella (LK Jones) Boerema (formerly Ascochyta pinodella LK Jones) did not produce ascochitine. Crystalline ascochitine was found to be of moderate toxicity to Artemia salina larvae (LC50 = 85 μg/cm3 brine shrimp medium) Extracts of Phoma medicaginis var. pinodella cultures were found to be highly toxic to Artemia salina. Addresses: 1, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University, 02-766 Warszawa, Poland.

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Biosynthesis and Preparation of Five Alternaria Metabolites 1

1

M Kostecki , J Grabarkiewicz-Szczsna , and J Chelkowski

2

Mycotoxin Research 1991, Vol. 7 (1), 3-7

Metabolites of Alternaria alternata were produced on rice as a solid substrate, chosen out of 6 substrates as the most useful. Optimal methods of extraction, purification, and separation of 5 metabolites were elaborated, using liquid - liquid partition, column chromatography, and preparative TLC. Alternariol, alternariol methyl ether, and copper salt of tenuazonic acid were obtained as crystals, altertoxin and altenuene as a film. Addresses: 1, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University, 02-766 Warsaw, Poland.

Regulation of Mycotoxin Biosynthesis in Alternaria P Häggblom1 and M Hiltunen2 Mycotoxin Research 1991, Vol. 7 (1), 7-10

The genus Alternaria is responsible for different plant diseases such as tobacco brown spot, tomato blight, and citrus seedling chlorosis but can also be present during storage of grain. The objective of the present paper is to summarize the knowledge concerning regulation of secondary metabolism in Alternaria, particulary A. alternata (A. tenuis). The paper mainly deals with regulation of polyketide biosynthesis, one of the major pathways leading to the biosynthesis of mycotoxins in Alternaria. The mostly studied Alternaria mycotoxins are dibenzopyrones such as alternariol (AOH) and alternariol monomethyl ether (AME) and altenuene along with the tetramic acid tenuazonic acid. The biosynthesis of Alternaria mycotoxins has been reviewed by Stinson. Most information is available for the biosynthesis of the polyketides AOH / AME while a few biosynthetic studies have been accomplished for tenuazonic acid. Addresses: 1, National Veterinary Institute, Box 7073, S-750 07 Uppsala, Sweden; 2, Dept of Plant Physiology, Uppsala University, Box 540, S-751 21 Uppsala, Sweden.

Toxicity of Mycotoxins Produced by Four Alternaria Species to Artemia salina Larvae P Zajkowski1, J Grabarkiewicz-Szcesna2, and R Schmidt3 Mycotoxin Research 1991, Vol. 7 (1), 11-15

22 isolates of Alternaria alternata, A. raphani, A. consortiale, and A. chartarum were examined for the production of alternariol (AOH), alternariol methyl ether (AME), altenuene (ALT), altertoxin I (ATX I), and tenuazonic acid (TA) on wheat grain and for toxicity of culture extracts to Artemia salina larvae. The total amount of 5 toxins produced under laboratory conditions ranged from 5 mg/kg to 11.112 mg/kg. The toxic extracts showed EC50 values in the range of 3.3 to 144.5 mg/mL. There was no correlation between toxicity of extracts to Artemia salina and the amount of mentioned mycotoxins in culture. Adresses: 1, Dept of Plant Pathology, Agricultural University, P-02766 Warsaw, Poland; 2, Dept of Chemistry, Agricultural University, P-60625 Poznan, Poland; 3, Mycotoxin Research Laboratory, D-6500 Mainz.

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The Occurrence of Fusarium and Alternaria on Faba Bean Seed E Zakrzewska Mycotoxin Research 1991, Vol. 7 (1), 16-18

The studies on the occurrence of Fusarium and Alternaria in faba bean seed have been conducted for three years. The fungal flora in seed lots from healthy plant and from plant inoculated with Ascochyta fabae were determined. In addition the occurrence of Fusarium and Alternaria in three parts of seed: testa, cotyledones, and embryo axis were investigated. It was found that Fusarium and Alternaria were present in faba bean seed, but the frequency of occurrence varied depending upon cultivar and experimental year. Address: Plant Breeding and Acclimatization Institute, Radzikow, 05-870 Blonie, Poland.

Effects of T-2 Toxin and Its Congeners on Membrane Functions of Cultured Human Fibroblasts Y-W Kim, RP Sharma, and Y Elsner Mycotoxin Research 1991, Vol. 7 (1), 19-28

Cytotoxicity of T-2 toxin, HT-2 toxin, acetyl T-2, neosolaniol, and T-2 tetraol was compared between normal human fibroblasts and mutant I-cell human fibroblasts, which only produce 10 to 15 % of lysosomal hydrolases present in normal fibroblasts. Both cleavage of 3-(4,5-dimethyl-2-thiazolyl)-2,5diphenyl-2H-tetrazolium bromide (MTT) and cell count by hemocytometer were used for evaluations. For all toxins, dose - related effects on both types of cultures were evident. Cytotoxicity of the above mycotoxins on both cell lines were similar, indicating that lysosomal enzymes were not involved in the toxicity of T-2 toxin and its congeners. An inhibitor of lysosomal cysteine proteases (E-64) did not alter the cytotoxicity of T-2 toxin. The decreasing order of toxicity was T-2 toxin, HT-2 toxin, neosolaniol, acetyl T-2 toxin, and T-2 tetraol in both cell lines. When normal human fibroblasts were loaded with the fluorescent dye Lucifer yellow CH (LY), a subsequent treatment of T-2 toxin did not disrupt lysosomal membranes. The uptake of LY was not affected by T-2 toxin, which indicated that T-2 toxin did not interfere with the endocytic pathway. Results indicate that T-2 toxin and its congeners do not exert their primary toxic effect through lysosomal enzymes, membranes, or via the endocytic pathway. Address: Toxicology Program, Utah State University, Logan, UT 84322-5600, USA.

Investigations on Possible Genotoxic Effects of Fusarium Toxins in Boars 1 1 2 1 2 K Lusky , U Wagner , B Stähr , K-D Doberschütz , and W Peter

Mycotoxin Research 1991, Vol. 7 (1), 29-34

For several weeks, boars were fed feedstuff containing mycotoxins (zearalenone, nivalenol, and deoxynivalenol). To determine a possible mutagenic effect of this feedstuff, the boars were examinated for structural chromosome aberrations in the lymphocytes. The investigations indicate a genotoxic effect on the boars´ lymphocytes. Addresses: 1, Institut für Veterinär - Pharmakologie und Toxikologie, O-1280 Bernau, Germany; 2, Institut für Biotechnik der Fortpflanzung, O-1282 Schönow, Germany.

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Survey of Total Aflatoxins in Camels Sera by Enzyme Linked Immunosorbent Assay (ELISA) 1

NA Osman and F Abdel-Gadir

Mycotoxin Research 1991, Vol. 7 (1), 35-38

A survey of total aflatoxins was carried out on samples of camels blood sera in order to assess aflatoxin hazards in camels following some deaths among animals during July through September, 1990 in Al Ain area, about 16 km from Abu Dhabi City. Epidemiological and laboratory based studies indicated that the outbreak was associated with the consumption of mould damaged feed. A. flavus and A. parasiticus and varying amount of aflatoxin were detected in samples testet. 53 samples of camel sera were collected for the analysis: 16 out of 53 samples were collected from apparently clinically healthy racing camels. All these samples were examined for total aflatoxins. All of the 16 samples were found to contain aflatoxins ranging from 5 to 50 ng/mL total aflatoxin and 28 out of the 37 samples were found to contain aflatoxin at the range of 2 to 12 pg/mL total aflatoxins. Address: Veterinary Laboratory, Agriculture Department and Animal Production, PO Box 1004 Alain, Abu Dhabi, United Arab Emirates.

Mutagenicity of Potentially Carcinogenic Mycotoxins Produced by Fusarium moniliforme WCA Gelderblom and SD Snyman Mycotoxin Research 1991, Vol. 7 (2), 46-52

The mutagenic behaviour of two potentially carcinogenic mycotoxins produced by Fusarium moniliforme was investigated in the Salmonella mutagenicity test using tester strains TA97a, TA98, TA100, and TA102. The mutagenic response obtained with fusarin C (1, 5, and 10μg/plate) against tester strains TA98 and TA100 in the presence of microsomal activation confirmed previous observations on the mutagenic behaviour of this mutagen while that obtained against TA97a is reported for the first time. No dose - response relationship could be detected for the concentration levels (0.2, 0.5, 1, 5, 10 mg/plate) tested for FB1, FB2, and FB3 against any of the tester strains used in either the plate incorporation and / or the pre-incubation test. A cytotoxic effect was obtained at concentration levels of 5 and l0 mg/plate in the absence of the microsomal activation mixture. From the studies it became evident that F. moniliforme produces two compounds, a mutagenic compound, fusarin C which has been shown to lack carcinogenic activity in rats and the non- mutagenic fumonisin B mycotoxins of which FB1 is known to be responsible for the hepatocarcinogenicity of the fungus in rats. Address: Research Institute for Nutritional Diseases, PO Box 19070, Tygerberg 7505, Republic of South Africa.

Production of Zearalenone, Nivalenol, Moniliformin, and Wortmannin from Toxigenic Cultures of Fusarium Obtained from Pasture Soil Samples Collected in New Zealand 1 2 3 HK Abbas , CJ Mirocha and R Gunther

Mycotoxin Research 1991, Vol. 7 (2), 53-60

One culture of F. avenaceum, 4 cultures of F. oxysporum, and 11 cultures of F. sambucinum were isolated from soil samples of pasture in New Zealand in 1987. All cultures, when grown on rice media and fed to rats caused a weight loss in rats as well as toxic signs including hemorrhaging and congestion, uterine enlargement, and hematuria. 6 out of 16 cultures caused death in rat feeding tests. F. oxysporum # 1 killed rats (feeding test) within 5-l2 hrs. 10 cultures produced zearalenone (19 to 8,849 ppm), 8 cultures produced nivalenol (32 to 117 ppm), 1 culture, F. sambucinum # 8, produced wortmannin (40 ppm), and 5 cultures produced moniliformin (19 to 9,000 ppm). We report for the first 202

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1991

time the co-occurrence of zearalenone, nivalenol, and moniliformin produced by F. sambucinum # 3 in culture. F. avenaceum # 1 and F. oxysporum cultures (nos 1, 2, and 3) produced moniliformin alone. F. oxysporum # 4 produced zearalenone alone as well. F. sambucinum # 5 caused erythema in the small intestine of rats and 100% mortality and did not produce any known toxin(s). Nivalenol when administered to the stomach of rats orally at levels 10, 20, and 40 mg/kg body weight caused inflammation in the intestines, coma, and death. The mycotoxins T-2 toxin, HT-2 toxin, T-2 tetraol, diacetoxyscirpenol (DAS), monoacetoxyscirpenol (MAS), deoxynivalenol (DON), 3-acetyl-and 15acetyldeoxynivalenol, depoxynivalenol, fusarenon-X, alpha - and beta - zearalenone, and fusarochromanone (TDP-1) were not detected in the extracts of these cultures. Addresses: 1, USDA-ARS, Southern Weed Science Lab, Stoneville, MS 38776, USA; 2, Departments of Plant Pathology , and 3, Laboratory Medicine and Pathology, School of Medicine, University of Minnesota, St Paul, MN 55108, USA.

Inhibitory Effect of Hena (Lawsonia inermis Leaves) and Carrot Root on Aflatoxin Production by Aspergillus parasiticus EI-Sayed AM Abd Alla and A Badawey Mycotoxin Research 1991, Vol. 7 (2), 61-68

Experiments were undertaken to evaluate the effect of some natural products (hena, and carrot root) on growth and aflatoxins production by Aspergillus parasiticus FRR 2752. Powdered hena (0.5 and 5%) inhibited mycelial growth and delayed 1 sporulation of A. parasiticus during 7 days. The inhibition of growth was increased with increasing the added amount. Aflatoxins production by A. parasiticus was reduced with 40-100% in the presence of hena (Lawsonia inermis leaves). Carrot root extract stimulated the fungal growth and aflatoxin production, whereas carrot root fibers slightly enriched fungal growth, inhibited aflatoxins production (B1, G1 and G2), but there was no inhibition of aflatoxin B2 production by A. parasiticus. Address: Mycotoxins Central Lab, National Research Centre, Dokki, Cairo, Egypt.

Natural Occurrence of Zearalenone in Rice and Soybean Produced in Korea Lee Y-W1, Kim J-G1, Chung D-H2, Roh P-U3, and Pestka JJ4 Mycotoxin Research 1991, Vol. 7 (2), 69-72

88 rice and 75 soybean samples were collected from 8 provinces of Korea from March through September in 1988. The Fusarium mycotoxins, zearalenone was analyzed by direct competitive enzyme linked immunosorbent assay. 10.2% of rice and 9.3% of soybean samples contained detectable zearalenone. The average levels of zearalenone of rice and soybean samples were 11.78 μg/kg and 7.70 μg/kg, respectively. Addresses: 1, Graduate School of Public Health, Seoul National University, 28, Yunkeun- Dong, Chongro-Ku, Seoul 110-460, Korea; 2, Gyeongsang National University, Chinju 660 -701, Korea; 3, Bergen County Health Department, Paramus 07652, NJ, USA; 4, Michigan State University, East Lansing, MI 48824, USA.

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Myrotoxins From a Plant Pathogenic Isolate of Myrothecium roridum BB Jarvis, FT Comezoglu, S Wang, and HL Ammon Mycotoxin Research 1991, Vol. 7 (2), 73-78

An isolate of M. roridum (ATCC 52485) which is a potent pathogen to muskmelon was shown to produce a series of macrocyclic trichothecenes, the myrotoxins, heretofore found to be produced only by an isolate of M. roridum which is a pathogen to tomato. The fact that these two isolates are virulent pathogens and both produce the same potent mycotoxins suggests that these novel trichothecenes may play an important role in the pathogenicity of the fungi. Address: Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742, USA.

Screening for Resistance to Aspergillus flavus and Aflatoxin Production in Groundnut S Desai, MP Ghewande, G Nagaraj, P Narayan, S Chauhan, and H Singh Mycotoxin Research 1991, Vol. 7 (2), 79-84

All the varieties, advanced breeding lines, germplasm lines, and wild species used in the experiments differed significantly for their ability to allow invasion and aflatoxin production by an aflatoxigenic Aspergillus flavus strain. lnfection and colonisation were strongly correlated (r = 0.82), while there was no relation between infection and aflatoxin content or colonisation and aflatoxin content (r = 0.15). The varieties ICGS11 and S206 supported less infection and colonisation (range 35 to 40%). Lowest aflatoxin content was recorded in Chitra (3,200 ppb), while it was highest in Kaushal (38,250 ppb). A cross derivative of GAUG 1 x NCAc 17133 RF showed lowest infection and colonisation (86,3 and 25,28%, respectively), and also supported moderate aflatoxin production (4,000 ppb). Among germplasm lines spancross supported lowest aflatoxin production (2,026 ppb) while both the wild species vz. ICG 8127 and ICG 8128 were highly susceptible to infection, colonisation, and aflatoxin production. Address: National Research Centre for Groundnut (ICAR), Junagadh-362 015, Gujarat, India.

Aflatoxin and Ochratoxin - A Contamination of Dried Figs (Ficus carina L) from the 1988 Crop Özay G and Alperden I Mycotoxin Research 1991, Vol. 7 (2), 85-91

The study examines the occurrence of aflatoxin and ochratoxin A in the 1988 dried figs crop. Mycotoxin content, moisture, and aw (water activity) were analyzed in a total of 103 fig samples collected from various orchards and different stages of fig processing. Aflatoxins (B1, B2, G1, and G2) were present in 29% of the samples examined at 0.5-63.0, 0.5-37.7, 0.5-78.3, and 0.5-12.5 μg/kg, respectively. Ochratoxin A was detected in only 3% of the samples at 5.2 - 8.3 μg/kg. The moisture (and aw) values of the fruits were found suitable for mycotoxin formation in firm ripened and shrivelled figs. Address: TÜBITAK, Marmara Research Center, Department of Food and Refrigeration Technology, PO Box 21, 41401 Gebze-Kocaeli, Turkey.

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Identification of Chlamydosporol, a Mycotoxin Isolated from a Culture of Fusarium tricinctum Michele Solfrizzo and Angelo Visconti Mycotoxin Research 1991, Vol. 7A (Part 1), 2-7

Fusarium tricinctum strain (KF 260) isolated from wheat in Poland, produced on maize cultures a compound (C11H14O5, MW 226) toxic to Artemia salina. The compound, identified by various spectroscopical techniques as 5-hydroxy-4-methoxy-6‚8a-dimethyl-6,7-dihydro-2H,8aH-pyrano/2,3b/pyran-2-one, was identical to chlamydosporol, a mycotoxin recently discovered in cultures of F. chlamydosporum isolated from rice. The compound showed inhibitory effect on human lymphocyte proliferation at concentration af 25 μg/mL. LD50 on A. salina was 56 μg/mL. Address: Istituto Tossine e Micotossine da Parassiti Vegetali, C.N.R., Via Amendola 197/F, 70126 Bari, Italy. Influence of Dietary Fibre on Plasma and Urinary Levels of Zearalenone and Metabolites in Swine 1 2 2 3 M Olsen , K Malmlöf , H Pettersson , and J Grajewski

Mycotoxin Research 1991, Vol. 7A (Part 1), 8-11

The effect of dietary fibre on zearalenone absorption, metabolic pathway and urinary excretion was investigated in a prepubertal gilt. Levels of zearalenone and metabolites were measured in the portal vein, in the carotid artery and in urine after feeding one portion of a standard diet or a high fibre diet (85 % standard diet + 15 % oat husks), both containing 5 ppm zearalenone. Independently of diet, the levels of zearalenone in the portal vein were higher than in the carotid artery during the following 48 hrs. Regarding the metabolite -zearalenol, strikingly lower systemic levels were observed when high fibre diet was fed to the animal. The decrease of -zearalenol level, due to high fibre diet, was also reflected in urine. The total amount of zearalenone and -zearalenol excreted in urine, during 24 hours after feeding, was about 9 % of the zearalenone intake, independently of diet. Almost all zearalenone and -zearalenol were found conjugated with glucuronic acid in both plasma and urine. Addresses: 1, National Food Administration, Biology section, S-751 26 Uppsala, Sweden; 2, University of Agicultural Sciences, Dept. of Animal Nutrition and Management, S-750 07 Uppsala, Sweden; 3, Academy of Agricultural Sciences and Technology, 85-029 Bydgozcz, Poland. Histopathological and Cytological Abnormalities Induced by Fusarial Toxins in Mice (Mus musculus) KS Bilgrami Mycotoxin Research 1991, Vol. 7A (Part 1), 12-16

Three fusarial toxins viz.‚ zearalenone (ZEA), deoxynivalenal (DON) and T-2 were administered orally to mice (Mus musculus) under two different treatments. In the first treatment, the toxins were fed at the rate of 0.5 mg/kg body weight (ZEA), 3 mg/kg body weight (DON), and 0.5 mg/kg body weight (T-2) for eight weeks (twice a week). In another set of experiments the toxins were given daily by mixing with the feed at a concentration commonly found in food and feed items as natural contaminants. The affected animals showed significant changes in body weight. T-2 toxins caused swelling of stomach in most of the treated animals. Portal and peripartal fibrosis in liver was also very common. Enlargement of urinary bladder was observed in ZEA fed animals. Cytological studies in bonemarrow cells showed fragmentation, chromosome breaks, stickiness and clumping of chromosome. Address: University Department of Botany, Bhagalpur University, Bhagalpur - 812 007, India. 205

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Broiler Chickens Fusariotoxicosis - A Histopathological Study of Lymphoid Organs and Testes 1 1 2 2 M Katkiewicz , K Czuminska , J Chelkowski , and N Rakowska

Mycotoxin Research 1991, Vol. 7A (Part 1), 17-25

The addition of five Fusarium species, cultured on wheat grain, to the standart chicken diet DKA starter, caused atrophic changes in the thymus and testes, as observed in the microscopic picture of these organs. The degree of lesions were depended on the Fusarium species and its amount added to the standart diet. Addresses: 1, Departament of Pathology, Veterinary Faculty, Warsaw, ul. Grochowska 272, Poland; 2, Departament of Phytopathology, Agricultural University of Warsaw, Poland.

Nivalenol Production by Fusarium poae Hans Pettersson Mycotoxin Research 1991, Vol. 7A (Part 1), 26-30

Fusarium sp. were isolated from Swedish nivalenol containing grain and tested for toxin production. Only F. poae, 6 of 10 isolates, produced nivalenol. Highest production (44.7 μg/g) was obtained cultured on rice during 4 week at room temperature and under near UV-light. Five F. poae isolates from other countries did not produce nivalenol but T-2/HT-2 toxin. One Swedish isolate produced both types of trichothecenes. Treatment with fungicides in a F. poae infected experimental field reduced the nivalenol concentration in the harvested grain. Address: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, P.B. 7024, S-750 07 Uppsala, Sweden.

Diacetoxyscirpenol Production by Fusarium sambucinum strain KF 735 on Solid and Liquid Media 1 2 3 4 J Perkowski ‚ D Latus-Zitkiewicz ‚ PB Hamilton , and J Chelkowski

Mycotoxin Research 1991, Vol. 7A (Part 1), 31-37

The yield of diacetoxyscirpenol (DAS) production by F. sambucinum strain No KF 735, isolated from potato tuber with dry rot symptoms, cultured on solid media and on liquid medium, has been examined. The amount of DAS produced within 28 days at 25 °C in the cultures grown on solid media (wheat, rye, rice, oats, corn, barley, triticale and malt) reached 238 mg/kg r 9 to 789 r l6 mg/kg (mean r standard error; n=3), on potato cubes  55 r 3 mg/kg and on the potato extract  147 r 5 mg/dcm3. The best substrates for crystalline compound production were malt and barley grain. Addresses: 1, Agricultural University of Pozna, Department of Chemistry, ul. Wojska Polskiego 28, 60-624 Pozna, Poland; 2, Agricultural University of Pozna, Institute of Food Technology of Plant Origin, ul. Wojska Polskiego 31, 60—624 Pozna, Poland; 3, Department of Poultry Science, North Carolina State University, Releigh, North Carolina 27695-7608, USA; 4, Agricultural University of Warsaw, Department of Plant Pathology, ul. Nowoursynowska 133, 02-766 Warsaw, Poland.

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Genetic Analysis as a Tool for Prediction of Mycotoxin-Producing Fusarium Strains 1

2

1

Logrieco A , Bottalico A ‚ and Mulè G

Mycotoxin Research 1991, Vol. 7A (Part 1), 38-42

The formation of fertile crosses (when possible), nuclear DNA relatedness and sequence comparison of select region of a large subunit (25S) ribosomal RNA are suitable methods for taxonomic determination and a phylogenetic classification of the strains. These genetic methods can also be used to predict which Fusarium strains produce mycotoxins since the genetic placement of the strains has been shown to have a good correlation with mycotoxin production ability at the species level. Addresses: 1, Istituto tossine e micotossine da parassiti vegetali del Consiglio Nazionale delle Ricerche, 70125 Bari, Italy; 2, Dipartmento di Patologia vegetale dell‘Università degli Studi, 70126 Bari, Italy.

Analysis of Morphometrical Features of Macroconidia from Fusarium spp. Hagen, B1, Hagen, C2 Mycotoxin Research 1991, Vol. 7A (Part 1), 43-49

No abstract available. Addresses: 1, Friedrich-Schiller-Universität, Biolog.-Pharm. Fakultät, Allgemeine Mikrobiologie, Neugasse 24, Jena 6900, Germany; 2, Friedrich-Schiller-Universität, Biolog.-Phar. Fakultät, Institut für Allgemeine Botanik, Thomas Mann-Str. 44, Jena 6900, Germany.

FUSKEY, an Interactive Computer Key to Common Fusarium Species Ulf Thrane Mycotoxin Research 1991, Vol. 7A (Part 1), 50-53

An interactive computer key to 17 common Fusarium species has been developed. The key, FUSKEY, uses morphological, physiological and chemical criteria and is running on a PC (MS-DOS structured) using easy obtainable software. Address: Department of Biotechnology, Building 221, The Technical University, DK-2800 Lyngby, Denmark.

The Similarity of Antigenic Proteins of Fusarium nivale (Fr.) Ces Isolates Marian Wiwart Mycotoxin Research 1991, Vol. 7A (Part 1), 54-57

The degree of similarity among dissolvable in water proteins obtained from 10 Fusarium nivale isolates was studied. Tandem crossed immunoelectrophoresis method was applied. Two immunological antisera: anti-F. nivale 4 and anti-F. nivale 15 were used in the investigation. Their homologous reactions resulted in 13 and 12 precipitation bands, respectively. The results allow to divide the isolates into three groups, according to their protein composition. Address: Department of Plant Breeding and Seed Production, Agricultural and Technical University, Olsztyn, Poland.

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Ecology and Taxonomy of Fusarium Species in Poland 1

2

H Kwana and J Chelkowski

Mycotoxin Research 1991, Vol. 7A (Part 1), 58-63

The paper presents 21 Fusarium species occurring in Poland on the field crops (mostly on cereals, maize, potato and Papilionaceae plants), woody plants, grasses, vegetables and ornamentals as well as on kernels or seeds of these hosts and soils. Additionally the commonly observed symptoms on abovementioned plants and the informations about regions of the highest disease occurrence are added. However the paper results mainly from authors’ investigation on the Fusarium species occurrence in Poland, it encloses also the available, post-war literature data on the fusariosis in Poland in the past. Majority of Fusarium species cited had been identified according to Nelson et al. taxonomic system. Comparative listing of the synonyms within eleven Fusarium sections as used in three monographs is presented. Addresses: 1, Department of Forest Pathology, Agricultural University, 60-625 Pozna, ul. Wojska Polskiego 71 c, Poland; 2, Department of Plant Pathology, SGGW-AR, 02-766 Warsaw, ul. Nowoursynowska 166, Poland.

Resistance Level and Toxin Contamination in Wheat and Corn after Artificial Inoculation 1 1 2 3 Mesterházy Á , Bartók T , Téren J , and Korom Á

Mycotoxin Research 1991, Vol. 7A (Part 1), 64-67

Most of the work made until now in toxicology of cereals did not consider enough the problem of resistance level and toxin contamination. A preliminary condition for such work is the possible correct measurement of resistance. As in earlier phases of the program this problem has been solved, we could begin the investigation of this relationship with the assumption that a high level of resistance will result in a low toxin contamination level. Addresses: 1, Cereal Research Institute, Szeged, Hungary; 2, Animal Health Service Laboratory, Szeged, Hungary; 3, Service Laboratory of State Farms, Szeged, Hungary.

Resistance Components of Wheat to Scab Á Mesterházy Mycotoxin Research 1991, Vol. 7A (Part 1), 68-70

No abstract available. Address: Cereal Research Institute, Szeged, Hungary.

Moniliformin and the European Corn Borer (Ostrinia nubilalis) H Lew, A Adler, and W Edinger Mycotoxin Research 1991, Vol. 7A (Part 1), 71-76

A representative survey was made of maize ears of the 1988 and 1989 crop in Austria to establish the influence of corn borer injuries on Fusarium species involved in ear fusariosis and Fusarium toxin production. The Fusarium species most frequently isolated from rot-damaged ears were F. sacchari var. subglutinans (about 50 %) and F. graminearum (about 30 %). There was a striking difference between the Fusarium species of the Liseola and the Discolor section concerning their occurrence on 208

Mycotoxin Research Vol. 24, No. 4 (2008), Abstracts 1991

corn borer-damaged ears. More than 80 % of the ears infected with F. sacchari var. subglutinans and F. verticillioides, but less than 15 % of the ears infected with F. graminearum, F. crookwellense and F. culmorum showed corn borer injuries. Toxin analyses of the infected ears corresponded to the known toxigenicity of the respective Fusarium species. Ears infected with F. sacchari var. subglutinans contained moniliformin (up to 20 mg/kg), those infected with F. verticillioides fumonisin B1 and B2 (up to 15 mg/kg). In ears infected with F. graminearum, F. culmorum and F. crookwellense zearalenone (up to 40 mg/kg) and deoxynivalenol (up to 500 mg/kg) or nivalenol (up to 10 mg/kg), respectively, could be detected. Hence measures to combat the European corn borer will mainly reduce moniliformin and fumonisin contamination, but will affect zearalenone, deoxynivalenol and nivalenol contents of the ears to a much lesser extent. Address: Federal Institute of Agrobiology Linz, Austria.

Cultural Characteristics and Zearalenone Production by Strains of Fusarium from Wheat Antónia Šrobárová and Marta Bukováková Mycotoxin Research 1991, Vol. 7A (Part 1), 77-83

Conidia of the species Fusarium culmorum (W.G.Sm.) Sacc. and Fusarium graminearum Schwabe are characterized by variability in zearalenone production and dimensions depending on the substrate. The sporulation of isolates from some wheat cultivars have been deprived in vivo and in vitro in the first passage, but not their pathogenicity and toxic metabolites production. Nonsporulating strains produced lower quantites of zearalenone than sporulating ones. Liquid filtrates of such nonsporulating strains had a high phytotoxic effect on wheat caryopses. The crystalline toxin diacetoxyscirpenol (0.25 μg/ml) had low phytotoxic effect on wheat caryopses. Address: Institute of Experimental Phytopathology and Entomology, Slovak Academy of Science, Ivanka pri Dunaji, Czechoslovakia.

Zearalenone Production in Wheat Cultivars Infected With the Fungus Fusarium graminearum Schwabe Marta Bukováková, Antónia Šrobárová, and Igor Vozár Mycotoxin Research 1991, Vol. 7A (Part 1), 84-90

In wheat plants of the cultivars „Danubia“, „Agra“, „Selekta“ and „Jubilejna“ the fungus Fusarium graminearum Schwabe produced toxic metabolite zearalenone (F-2) which simultaneously influenced the development of plants characterized by a lower germinating capacity, a reduced growth rate and a higher production of side branches. The presence of Fusarium graminearum was confirmed only in infected plants after plating of organs (root, stem base, stem) and soil on agar medium. The mycotoxin production is dependent on the pathogen development in host plants. The F-2 level progressed from the root into the soil, stem base and stem. The highest F-2 production was identified in cultivar “Selekta“, the lowest in cultivar “Danubia“. The highest F-2 level (in all wheat cultivars) was identified in the stem base. Address: Institute of Experimental Phytopathology and Entomology, Slovak Academy of Science, Ivanka pri Dunaji, Czechoslovakia.

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Susceptibility of Selected Winter Wheat Cultivars Produced in Poland to Fusarium Head Blight 1 2 3 Wakuliski W , Solfrizzo M , and Perkowski J

Mycotoxin Research 1991, Vol. 7A (Part 1), 91-96

The susceptibility of six winter wheat cultivars to Fusarium head blight has been studied. The lowest infected plants intensity, mean degree of head damage and fusariosis index exhibited cultivar Grana which also cumulated the lowest amount of trichothecenes (deoxynivalenol and derivatives). Possibility to produce nivalenol by Polish strains has been found in F. graminearum. Addresses: 1, Department of Plant Pathology, Agricultural University, 02-766 Warsaw, Poland; 2, Istituto Tossine e Micotossine da Parassiti Vegetali, CNR, Bari, Italy; 3, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland.

Pathogenicity of Fusarium spp. Contributing to the Stalk Rot of Maize in Poland M Proczuk, S Proczuk, and M Messyasz Mycotoxin Research 1991, Vol. 7A (Part 2), 97-101

During 1985-1989 a stalk rot of early maturity hybrids of maize was studied in Radzikow (Central Poland). It was found that Fusarium species were dominant on plants with stalk rot symptoms. Spectrum of Fusarium spp. had changed within the years. The most frequently isolated were: F. subglutinans, F. culmorum and F.crookwellense. When the disease developed early in the season, F. graminearum was also present. Predominant species were examined for their pathogenicity according to the modified method of Molot, Simone (1967). Isolates of F. graminearum and F. culmorum were found to be strong pathogens, F. crookwellense and F. subglutinans  moderate, and F. oxysporum and F. equiseti were the weak ones. Address: Plant Breeding and Acclimatization Institute, Radzikow 05870 Blonie, Poland.

A Study of the Correlations Between the Amount of Deoxynivalenol in Grain of Wheat and Triticale and Percentage of Fusarium Damaged Kernels J Perkowski1, J Chelkowski2, P Blaczak3, CHA Snijders4, W. Wakuliki2 Mycotoxin Research 1991, Vol. 7A (Part 2), 102-114

The correlation between the amount of deoxynivalenol (DON) and the percentage of Fusarium damaged kernels (FUK) in samples of wheat and triticale was studied. Samples of natural infected wheat grain, collected in 1986, 1987 and 1988 and of triticale collected in 1986 were used. Additionally, artificially inoculated wheat samples (10 genotypes inoculated with 3 F. culmorum strains of weak, medium and severe pathogenicity and samples of 10 triticale genotypes inoculated with F. culmorum and F. graminearum) were studied. Using statistical methods (the variance analysis, method of least significant difference (LSD), orthogonal contrast (OC) and minimum within groups sum of squares criterion (MSSC)), the samples were divided into two groups with respect to the attribute DON/FDK. To the first group belong samples of wheat and triticale, of which the heads were artificially inoculated with severely pathogenic strains of F. culmorum. In the samples of this group the amount of DON in kernels damaged with Fusarium increased by 0.46 mg/kg per 1 % of FDK. In the second group, consisting of naturally infected samples and samples from artificially inoculated heads the amount of DON increased 0.30 mg DON/kg per 1 % of FDK. The equation for the calculation of approximated amount of DON in farm and commercial lots of wheat and triticale after examination of percentage of FDK is given.

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Addresses: 1, Academy of Agriculture, Department of Chemistry, 60625 Pozna, Poland; 2, Agricultural University, Department of Plant Pathology, 02766 Warsaw, Poland; 3, Academy of Agriculture, Department of Mathematical and Statistical Method, 60637 Pozna, Poland; 4, Centre for Plant Breeding Research, P.O.Box 16, 6700 AA Wageningen, The Netherlands.

Cumulation of Mycotoxins in Maize Cobs Infected with Fusarium graminearum 1 2 3 1 Juliusz Perkowski , Jerzy Chelkowski , Ronald D. Plattner , and Piotr Goliski

Mycotoxin Research 1991, Vol. 7A (Part 2), 115-120

Maize cobs with Fusarium ear rot were collected at 1986 season and five infected by Fusarium graminearum were analyzed for presence of trichothecenes and zearalenone. Collected material was subsampled for Fusarium damaged kernels and corresponding axial stems and healthy looking kernels. All investigated cobs contained deoxynivalenol (DON) (range 18.0131.5 mg/kg) and zearalenone (ZEA) (range 0.382.17 mg/kg), in four cobs 15-acetyl-deoxynivalenol (15-AcDON) (range 5.26.2 mg/kg) was present and two cobs besides three all metabolites contained 3-acetyl-deoxynivalenol (3-AcDON) (range 0.50.8 mg/kg). The average of individual toxins amount in axial stems: in mg/kg was equal to: DON  110.36, ZEA  4.57, 15-AcDON  16.66, and 3-AcDON  1.32. Fusarium damaged kernels contained in average the following amount (mg/kg) of: DON 77.00, ZEA 0.98, 15-AcDON 3.78 and 3-AcDON 0.06. Healthy looking kernels contained DON 1.96 mg/kg and ZEA 0.07 mg/kg only. Cooccurrence of 3-AcDON and 15-AcDON in two samples was an interesting finding. The amount of DON in total cob was highly correlated (r = 0.94) with percentage of Fusarium damaged kernels in given ear. Addresses: 1, Department of Chemistry, Agricultural University of Pozna, ul. Wojska Polskiego 75, 60625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University of Warsaw, ul. Nowoursynowska 166, 02766 Warsaw, Poland; 3, Northern Regional Research Center, ARS-SDA, 1815 North University Street, Peoria, Ill. 61604, USA.

Pathogenicity of Seed Transmitted Fusarium spp. to Triticale Seedlings Arseniuk E1, AL Scharen2, and HJ Czembor1 Mycotoxin Research 1991, Vol. 7A (Part 2), 121-127

In the conducted studies 13 species of Fusarium were isolated into pure culture from triticale seed. Their pathogenicity was assessed under laboratory and greenhouse conditions. Most of the species studied were highly pathogenic to the first leaf seedlings of triticale „Grado“ and „Lasko“ under both sets of conditions. lt was shown, that seed-transmitted Fusarium spp. considerably reduced the ability of seeds to germinate and incited seedling blight. On average, triticale „Lasko“ was more resistant to Fusarium spp. than „Grado“, but in some instances a reverse reaction was observed. Addresses: 1, Plant Breeding and Acclimatization Institute, Radzikow, 05870 Blonie, Poland; 2, USDA  ARS, Department of Plant Pathology, Montana State University, Bozeman, MT 59717  0002, USA.

Fusariosis of Spring Barley Cultivated in Lublin Region Barbara Lacicowa, and Irena Kiecana Mycotoxin Research 1991, Vol. 7A (Part 2), 128-135

Diseases of spring barley in 19861988 seasons have been examined on barley plantations in Lublin region. Observations in eight weeks after sowing each year spring showed the occurrence of root rot and sheath rot in seedlings. As a result of mycological examination of infected seedlings 34 species of 211

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fungi were isolated: Fusarium spp. amounted up to 23% of all isolates. Each year, Fusarium culmorum and F. avenaceum were isolated, but F. graminearum only in 1987. On all inspected fields there occurred plants with eye-spots or necrotic stripes on lower internodes. As a result of fungi isolation the colonies belonging to 30 species were identified from stems and roots of examined plants. There was about 35% of fusaria between isolates each year. Fusarium culmorum was most frequently isolated. This fungus both from stems with two mentioned kinds of symptoms and from roots was isolated. Fusarium avenaceum each year and Fusarium graminearum in 1986 and 1988 were isolated. Mentioned there species were also isolated from kernels. Address: Department of Plant Pathology, Agricultural University, 20-934 Lublin, Poland.

Infection Ability of Mycelium and Spores of Microdochium nivale (Fr) Samuels Hallett to Lolium Perenne L. M Proczuk and M Messyasz Mycotoxin Research 1991, Vol. 7A (Part 2), 136-139

Mycelium and spores af Microdochium nivale (Syn. Fusarium nivale) were compared according to their ability to infect plants of Lolium perenne. The experiments were carried out according to the “cold chamber“ method (Cormack, Lebeau 1956 modified by Pronczuk 1987). Between these two types of inoculum significant differences were found. The spore inoculum did not give any symptoms while the mycelial inoculum incited a severe disease in plants of Lolium perenne during one month of incubation. Under laboratory conditions it was found that the spore cultures of Microdochium nivale grew very slowly at 01 °C, whereas their growth at 1820 °C was very fast. Growth of the mycelial cultures was not as profoundly affected by temperatures studied as the spore ones. It was concluded that to incite a disease the spore inoculum require longer incubation time than mycelial ones. The mycelial inoculum is more useful for screening of plants for resistance. Address: Plant Breeding and Acclimatization Institute Radzikow 05870 Blonie, Poland.

Toxinogenicity of Microdochium nivale (Fusarium nivale) Isolates from Cereals in Poland J Chelkowski1, P Golinski2, J Perkowski2, A Visconti3, M Rakowska4, and W Wakuliski1 Mycotoxin Research 1991, Vol. 7A (Part 2), 140-145

Microdochium nivale (Fusarium nivale) was found to be frequently occuring in Poland pathogen of small grain cereals heads, causing symptoms similar to those observed after infection of Fusarium species. In consecutive years since 1985 till 1989 the following percentage of wheat and rye ears infected with M. nivale and with Fusarium head blight symptoms was found: 34 %, 21 %, 42 %, 9 %, 46 % (wheat) and 57 %, 43 %, 65 %, 4 %, 47 % (rye) heads. However, in naturally infected rye and wheat samples (kernels and chaff), we did not detect toxins usually present in samples infected with fungi of genus Fusarium – such as deoxynivalenol and derivatives. Typical Fusarium trichothecene metabolites were also not present in cultures of 11 M. nivale strains, growing 35 weeks on rice (45 % water content) at 20 °C. Cultures of two typical isolates on wheat grain (strain KF 1124) and on rice (KF 245) were found to be non toxic to broiler chickens when present in amount 2040% in their diet. It can be concluded that M. nivale (F. nivale) representatives in Poland did not produce toxic metabolites neither under laboratory condition nor after cereal ears infection under field conditions. Addresses: 1, Department of Plant Pathology, Agricultural University of Warsaw, ul. Nowoursynowska 166, 02766 Warsaw, Poland; 2, Department of Chemistry, Agricultural University of Pozna, ul. Wojska Polskiego 75, 60625 Pozna, Poland; 3, Istituto Tossine e Micotossine da parasiti vegetali CNR, 70 126 Bari, Italy; 4, IHAR Radzikow k/Warszawy, Poland.

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Phytotoxicity of Deoxynivalenol to Wheat Calli I Menke-Milczarek and J Zimny Mycotoxin Research 1991, Vol. 7A (Part 2), 146-149

The main objective of this study was to determine the toxicity of DON (deoxynivalenol) in wheat tissue culture after two weeks of contact. It was observed that increase of toxin concentration in medium caused decrease of regeneration ability of the calli.Three tested genotypes showed different sensitvity to 25 ppm of DON. Address: Department of Tissue Culture, Plant Breeding and Acclimatization Institute, 00950 Warsaw, P.O. Box 1019, Poland.

Cytogenetic Changes in Plant Cells as Influenced by Mycotoxins Danuta Packa Mycotoxin Research 1991, Vol. 7A (Part 2), 150-155

The effect of mycotoxins deoxynivalenol (DON), diacetoxyscirpenol (DAS) and zearalenone (F-2) on actively dividing cells of cereals and field bean was studied. In general, mycotoxins resulted in a decrease of mitotic activity. However, the F-2 treatment increased the mitotic index in rye. DON appeared a strong inhibitor of cell divisions in wheat and field bean. DAS retarded divisions at the stage of methaphase and changes spiralization and colourability of metaphase chromosomes. The cytological picture of the DAS-treated cells indicates that this compound disturbs the functioning of the karyokinetic spindle leading to the formation of cells of multiplied number of chromosomes. Address: Department of Plant Breeding and Seed Production, University of Agriculture and Technology, Olsztyn, Poland.

Biosynthesis, Isolation, Purification and Separation of Zearalenone, Deoxynivalenol and 15-Acetyldeoxynivalenol 1 1 2 M Kostecki , P Goliski and J Chelkowski

Mycotoxin Research 1991, Vol. 7A (Part 2), 156-159

Fusarium graminearum KF–376 isolate was found to be able to form simultaneously three toxic metabolites: zearalenone (F-2), deoxynivalenol (DON) and 15–acetyldeoxynivalenol (l5–AcDON). Toxins were extracted with methanol–water 3: 1 (v/v) and liquid chromatography on charcoal-Kieselgel 60 column (preliminary) and Aluminiumoxid 90 column. Final separation of the metabolites was achieved on Kieselgel 60–Aluminiumoxid 90 column. Addresses: 1, Department of Chemistry, Agricultural University of Pozna, ul. Woiska Polskiego 75, 60625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University of Warsaw, ul. Nowoursynowska 166, 02–766 Warsaw, Poland.

Biosynthesis, Isolation, Purification and Separation of Nivalenol, Fusarenone-X and Zearalenone M Kostecki1, P Goliski1 and J Chelkowski2 Mycotoxin Research 1991, Vol. 7A (Part 2), 160-164

Fusarium graminearum KF 370 isolate is able to simultaneous biosynthesis of three toxic metabolites, namely: fusarenone-X (FUS), nivalenol (NIV) and zearalenone (F-2). After metabolites extraction 213

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with methanol–water (3:1) and defatting with n-heptane toxins were partitioned into chloroform layer. Purification of the compounds was performed on Celite 545–charcoal–Aluminiumoxid 90 column then metabolites were separated on Kieselgel 60 (200–300 mesh) column with developing solvent chloroform–methanol. This way FUS, NIV and F-2 were obtained as crystalline or high purity standards. Addresses: 1, Department of Chemistry, Agricultural University of Pozna, ul. Woijska Polskiego 75, 60625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University of Warsaw, ul. Nowoursynowska 166, 02–766 Warsaw, Poland.

Do the Pigments Differ Chemotaxonomically Fungi Belonging to Genus Fusarium? 1 2 2 1 P Goliski , J Chelkowski , P Zajkowski , and M Kostecki

Mycotoxin Research 1991, Vol. 7A (Part 2), 165-171

Biosynthesis of bikaverin – red pigment responsible for characteristic colour of Fusarium cultures and known as vacuolation and aging factor – by 60 Fusarium isolates belonging to 9 species was examined. We observed that bikaverin is a common metabolite produced by species of Liseola, Martiella, Elegans and Dlaminiella sections. Twenty-nine isolates of four Fusarium species (F. moniliforme, F. subglutinans, F. proliferatum and F. anthophilum), belonging to the following Liseola section were investigated or their ability to biosynthesis of secondary metabolites bikaverin, 8-methylfusarubin, 3,8-dimethylfusarubin, 8-methylbostrycoidin, nectriafurone and fusarin C. All examined isolates of section Liseola formed bikaverin and 8-methylfusarubin, while 21 isolates additionally produced 8methylbostrycoidin. Only F. moniliforme isolates were able to form fusarin C. We did not observe chemotaxonomical differences between Fusarium species according their ability to biosynthesis of such secondary metabolites as bikaverin, 8-methylfusarubin and 8-methylbostrycoidin. Addresses: 1, Department of Chemistry, Agricultural University of Pozna, ul. Woiska Polskiego 75, 60625 Pozna, Poland; 2, Department of Plant Pathology, Agricultural University of Warsaw, ul. Nowoursynowska 166, 02–766 Warsaw, Poland.

A Rapid Method for Extraction of Zearalenone and Zearalenols in Fermented Corn Ronald Vesonder1 and Piotr Golinski2 Mycotoxin Research 1991, Vol. 7A (Part 2), 172-177

A rapid extraction method is described for isolation of zearalenone and - and -trans-zearalenols from laboratory fermented corn. Corn fermented with Fusarium crookwellense at 25 °C for 2 weeks was agitated for 5 minutes in acetone. The acetone extract was evaporated to dryness and the remaining residue was chromatographed on a silica gel column with hexane:ethyl acetate (8:2). The products eluted with the hexane:ethyl acetate mixture in 1-1/2 column volumes and were identified by regular phase and C-18 reverse-phase thin layer chromatography. The products were verified by gas chromatography-mass spectroscopy. Product recoveries were 62.5–70 % for the zearalenols and 70 % for zearalenone in the range 0.5–50 mg/Kg. Addresses: 1, Northern Regional Research Center, USDA–ARS 1815 North University Street, Peoria, Il., 61604, USA; 2, Katedra Chemii Akademii Rolniczej w Poznaniu, ul. Wojska Polskiego 75, 60–625 Pozna, Poland.

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Contamination of Freshly Harvested Maize Kernels with Fusarium Mycotoxins in Bihar State, India KK Sinha Mycotoxin Research 1991, Vol. 7A (Part 2), 178-183

Freshly harvested maize samp1es, collected from different fields of Bhagalpur during January–March, 1989, were analysed for the presence of Fusarium species and their toxins. F. moniliforme was most common followed by F. roseum, F. sporotrichioides, F. graminearum and F. equiseti. Different strains of these species produced zearalenone (11.2–28.2 μg/g), DON (0.3–2.9 μg/g) and T-2 (5.2– 20.6 μg/g) toxins on mostrice medium. Fifteen percent, out of 86 maize samples analysed, were found to be contaminated with various levels of above toxins, which occurred either alone or in groups. Toxin concentration in contaminated samples varied from 0.76–1.5 μg/g (ZEN), 0.41–2.02 μg/g (DON) and 0.55–2.92 μg/g (T-2). Address: University Department of Botany, Bhagalpur University, Bhagalpur–812 007, India.

Fusariosis of Hop Ewa Solarska Mycotoxin Research 1991, Vol. 7A (Part 2), 184

No abstract available. Address: Laboratory of Hop Protection, Institute of Soil Science and Plant Cultivation, 21 002 Jastków, Poland.

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In Vitro Transformation of the Tremorgenic Mycotoxin Verruculogen 1

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A Bott , J Bauer , M Gareis , C Enders , B Kollarczik , and B Gedek Mycotoxin Research 1992, Vol. 8 (1), 2-8

A quantity of 50 mg of crystalline verruculogen was prepared from batch cultures of Aspergillus fumigatus for the use in the in vitro studies with S-9 liver fractions, feces suspensions, and cultures of Escherichia coli. Incubation of verruculogen with S-9 liver fractions from swine resulted in the transformation of the parent compound into TR-2 toxin and 3 other more polar. TR-2 toxin was also shown to be the main transformation product when incubating verruculogen with fecal suspensions or pure cultures of E. coli (O149:K88). Incubation times of more than 2 hours led to a complete degradation of verruculogen as well as TR-2 toxin. Addresses: 1, Institute for Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, University of Munich, Veterinaerstr. l3, W-8000 Munich 22, Germany; 2, Chair of Animal Hygiene, Technical University of Munich, Hohenbachernstr. 15, W-8050 Freising-Weihenstephan, Germany.

Alternaria alternata from Oilseed Rape: Mycotoxin Production, and Toxicity to Artemia salina Larvae and Rape Seedlings A Visconti, A Sibilia, and C Sabia Mycotoxin Research 1992, Vol. 8 (1), 9-16

A survey, carried out in Southern Italy on fungi colonizing oilseed rape (Brassicae napua L subsp oleifera DC) in the field, showed Alternaria alternata (Fries) Keissler as one of the predominant species. 11 strains of Alternaria alternata isolated from oilseed rape were cultured on rice to test the ability to produce mycotoxins. All strains produced mycotoxins, including tenuazonic acid (up to 12,000 mg/kg), alternariol (up to 200 mg/kg), alternariol monomethyl ether (up to 200 mg/kg), altertoxin-I (2 to 250 mg/kg), and altertoxin-II (2 to 70 mg/kg). Culture extracts containing tenuazonic acid were toxic to Artemia salina and inhibited seedling growth of oilseed rape. Little or no activity with the same bioassays was shown by culture extracts containing the remaining mycotoxins. No mycotoxins were detected in 16 samples of oilseed rape from processing factories. Address: Istituto Tossine e micotossine da parassiti vegetali, CNR, Viale Einaudi 51, I-70125 Bari, Italy.

Isolation of Cytochalasins A and B from Ascochyta lathyri 1 1 2 2 3 M Vurro , MC Zonno , A Evidente , R Capasso , and A Bottalico

Mycotoxin Research 1992, Vol. 8 (1), 17-20

The possible presence of toxic metabolites in the culture extracts of 24 Ascochyta strains, grown on autoclaved wheat, was ascertained by the use of the biological assay on brine shrimps (Artemia salina). Only in the culture extract of A lathyri, that showed a very high toxicity, the presence of cytochalasins A and B was revealed by tlc, 1H nmr and fab-mass spectra. Since A heteromorpha, previously described as the first Ascochyta species to produce cytochalasins, has been reclassified as Phoma exigua var heteromorpha, this is therefore the first report on the cytochalasin production by a true Ascochyta species. Addresses: 1, Istituto tossine e micotossine da parassiti vegetali del CNR, Via Einaudi, 51, I-70125 Bari, Italy; 2, Dipartimento di Scienze Chimico-agrarie dell´ Università „Federico II“ di Napoli, Via Università 100, I-80055 Portici, Italy; 3, Istituto di Patologia vegetale dell´Università degli Studi di Sassari, Via De Nicola, I-07100 Sassari, Italy. 216

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Aflatoxin Contamination in Field Mustard (Brassica juncea) Cultivars KS Bilgrami, AK Choudhary, and KS Ranjan Mycotoxin Research 1992, Vol. 8 (1), 21-26

7 varieties of mustards were cultivated in statistically designed field during 1988 through 1989 and 1989 through 1990 crops in order to ascertain the resistant or less susceptible varieties for aflatoxin elaboration. Under field condition, none of the varieties of mustard was found to be resistant. Maximum aflatoxin contamination was observed in the late planting time (15th November) crops. Insect (Lipaphis erysime) incidence had significant negative correlation (r = - 0.9354) with the yield (Q/ hectare) while it had significant positive correlation (r = 0.5705) with aflatoxin contamination. Address: University Department of Botany, Bhagalpur University, Bhagalpur - 812007, India.

Mycotoxins in Cereal Grain (Part 15). Distribution of Deoxynivalenol in Naturally Contaminated Wheat Kernels 1 2 J Chelkowski and J Perkowski

Mycotoxin Research 1992, Vol. 8 (1), 27-30

The analysis of deoxynivalenol (DON) in naturally infected wheat samples, after having been separated into four fractions through laboratory sieves, showed very low levels of DON in the fraction of largest kernels >2.8 mm (0 up to 1 mg/kg). The highest concentration of DON was found in fractions 2.2 to 2.5 mm and <2.2 mm with up to 14 mg/kg and l5 mg/kg DON, respectively. In two samples (fractions < 2.2 mm) nivalenol was detected in concentrations up to 1,4 mg/kg. Addresses: 1, Department of Plant Pathology, Agricultural University, 02-766 Warsaw, Poland; 2, Department of Chemistry, Agricultural University, 60-625 Pozna, Poland.

Decontamination of Fusarium Mycotoxins, Nivalenol, Deoxynivalenol, and Zearalenone, in Barley by the Polishing Process 1 1 2 3 U-S Lee , M-Y Lee , W-Y Park , and Y Ueno

Mycotoxin Research 1992, Vol. 8 (1), 31-36

Korean dehusked and unhusked barley naturally contaminated with Fusarium mycotoxins were polished using a Satake Grain Testing Mill. The pearled barley and bran fractions with different degrees of polishing were analyzed for nivalenol (NIV) and deoxynivalenol (DON) by gas chromatography with an electron capture detector, and for zearalenone (ZEN) by high - performance liquid chromatography with a fluorescence detector. NIV was detected in all the pearled barley fractions, but DON and ZEN were not detected in  27 % pearled barley fractions from dehusked barley and  36 % pearled barley fractions from unhusked barley. However, for all degrees of polishing, NIV, DON, and ZEN were detected in bran fractions. The levels of NIV, DON, and ZEN in the bran fractions increased several fold over the original barley. Polishing was effective in removing DON and ZEN from the naturally contaminated barley, but not NIV. Addresses: 1, Department of Food Technology, National Chung-ju Technical College, Jungwon-gun, Chung-buk 383-870, Korea; 2, Department of Pharmacognosy, College of Pharmacy, Chung-buk National University, Cheong-ju, Chung-buk 360-763, Korea; 3, Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Shinjuku - ku, Tokyo 162, Japan.

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Analysis of Mycotoxins in Cereals and Animal Feed Using a Multi-Toxin Gel Permeation Chromatography Clean-Up Method K Scudamore and M Hetmanski Mycotoxin Research 1992, Vol. 8 (1), 37-47

Gel permeation chromatography has been used to clean up extracts from cereals and animal feeds containing a range of mycotoxins. A mixture of dichloromethane: IM hydrochloric acid, 10:1 by volume is used as the extraction solvent and clean-up is carried out on a Bio - Beads S-X3 column using dichloromethane : ethyl acetate containing a small amount of formic acid as the elution solvent. Chromatographic separation and detection was by HPLC with fluorescence or ultraviolet detection although the choice of detection method is left to the user. The method has been tested for 14 mycotoxins and results are presented for cereals fortified with mycotoxins and for samples naturally contaminated with aflatoxins, citrinin, zearalenone and ochratoxin A. Address: Central Science Laboratory, Ministry of Agriculture, Fisheries and Food, London Road, Slough, Berkshire, SL3 7HJ, United Kingdom.

Fermentation of Wort Containing Deoxynivalenol and Zearalenone 1 1 2 2 Scott PM , Kanhere SR , Daley EF , and Farber JM

Mycotoxin Research 1992, Vol. 8 (2), 58-66

Wort containing deoxynivalenol and zearalenone, each added at a level of 1,9μg/mL, was fermented by 3 strains of Saccharomyces cerevisiae for 7 or 9 days to make beer. Analysis showed that deoxynivalenol was stable during this process. The major metabolite of zearalenone was -zearalenol, which formed in up to 69% of the initial zearalenone concentration, while up to 8.1% of the initial zearalenone was converted to -zearalenol. The major part of the metabolism of zearalenone occurred by 1 - 2 days. Control experiments. where the yeasts were omitted and deoxynivalenol, zearalenone and  - and  - zearalenol were added, showed good recovery and stability of the mycotoxins over the 7 - 9 day time period. No deoxynivalenol, zearalenone, -zearalenol or -zearalenol was detected in control yeast fermentations where they were not added to the wort. Addresses: 1, Food Research Division, Bureau of Chemical Safety, Food Directorate, Health Protection Branch, Health and Welfare Canada, Ottawa, Ontario, Canada K1AOL2; 2, Microbiology Research Division, Bureau of Microbial Hazards, Food Directorate, Health Protection Branch, Health and Welfare Canada, Ottawa, Ontario, Canada K1AOL2.

Evaluation of Test Kits for the Determination of Aflatoxins in Meat Products Mutti P, Dellapina G, and Spotti E Mycotoxin Research 1992, Vol. 8 (2), 67-72

A study was conducted to evaluate the performance of 4 different assays for rapidly screening samples of artificially contaminated hams and salami for the presence of aflatoxins (B1+B2+G1+G2) at concentrations  5μg/kg. Test samples were contaminated in the range of 0 - 100 μg/kg. At 0 μg/kg level no false positive (all <5 μg/kg) were found for all commodities by the kits tested; all test samples spiked at level > 20 μg/kg were found positive by each kit, while most of the errors associated in the assays occurred on samples containing <10 μg/kg. For samples either negative or contaminated above 20 μg/kg all the methods were suited for use as rapid screening tests. Address: Mycotoxin Department, Stazione Sperimentale per l‘lndustria delle Conserve Alimentari, Viale F Tanara, 31/A, I-43100 Parma, Italy.

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Bikaverin Production by Fusarium Species Chelkowski J, Zajkowski P, and Visconti A Mycotoxin Research 1992, Vol. 8 (2), 73-76

Production of bikaverin has been examined in 130 Fusarium isolates belonging to 21 species. The highest yield of bikaverin was produced on autoclaved rice-up to 2.5g/kg of dry culture. Bikaverin was produced by the following species: F. verticillioides, F. sacchari var. subglutinans, F. proliferatum, F. anthophilum, F. oxysporum F. dlamini, F. nygamai, F. napiforme, and F. solani. Species F. coeruleum, F. poae, F. sporotrichioides, F. tricinctum, F. chlamydosporum, F. culmorum, F. graminearum, F. cerealis (F. crookwellense), F. avenaceum, F. acuminatum, and F. equiseti did not produce bikaverin. The production of bikaverin determines the colour of the mentioned Fusarium species cultures on agar media and and on rice. The pigment has indicator properties and changes colour from red in acidic solution to violet-blue in alkaline. The role it plays in fungus metabolism is not elucidated. Address: Department of Plant Pathology, Warsaw Agricultural University, PL-02 766 Warsaw, Poland.

DNA Strand Break Induction, Mutagenicity, and Cytotoxicity of the Mycotoxins 11--Hydroxy-7-Deoxy- Rosenonolactone, Rosenonolactone, and Trichothecin Hengstler JG1‚ Löffler S1, Schaefer M3, Glatt HR1, Fuchs J1, Flesch P2, and Oesch F1 Mycotoxin Research 1992, Vol. 8 (2), 77-83

11  - hydroxy -7- deoxy - rosenonolactone (TSS1), a mycotoxin of the rosenane class, was tested on cytotoxicity, induction of DNA single strand breaks and mutagenicity. Its effects were compared to those of rosenonolactone and trichothecin. TSS1 had stronger antibiotic activity against Escherichia coli (EC 50: l0μg/mL) than rosenonolactone (EC 50: > 200μg/mL) but weaker activity than trichothecin (EC 50: 3μg/mL). The same order of activity was found for the inhibition of yeast fermentation (EC 50 of TSS1: 45μg/mL; EC 50 of rosenonolactone: >120 μg/mL; EC 50 of trichothecin: 3.4μg/mL). In the trypan blue exclusion test using V79 Chinese hamster cells, TSS1 proved to be cytotoxic (EC 50: 30μg/mL) at even lower doses than trichothecin (EC 50: 200μg/mL). Rosenonolactone had no significant toxicity up to the highest soluble concentration (500 μg/mL). DNA single strand breaks caused by TSS1 occurred at the same concentrations at which damage of the cell membrane became apparent. For trichothecin single strand breaks were detected only at concentrations at which the membrane was already highly damaged. No single strand breaks were observed in V79 cells after incubation with rosenonolactone up to the limit of solubility (500μg/mL). In the reversion assay with his - Salmonella Typhimurium strains TA 98 and TA 100, no mutagenicity was observed for any of the examined mycotoxins up to 800 μg/plate with and without the addition of a rat liver preparation for metabolism of the test compound. Addresses: 1, Institut für Toxikologie, Universität Mainz, Obere Zahlbacher Strasse 67, D-6500 Mainz, Germany; 2, Institut für Biochemie, Universität Mainz, Becherweg 39, D-6500 Mainz, Germany; 3, Bayer Diagnostic GmbH, Weissenseestrasse 101, D-8000 München 90, Germany.

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A Survey of the Occurrence of Fusarium Mycotoxins (Trichothecenes, Zearalenone, and Fusarochromanone) in Corn and Wheat Samples from Shaanxi and Shanxi Provinces, China 1 1 2 3 4 Luo Y , Yoshizawa T , Yang J-S , Zhang S-Y , and Zhang B-J

Mycotoxin Research 1992, Vol. 8 (2), 85-91

No abstract available. Addresses: 1, Department of Bioresource Science, Faculty of Agriculture, Kagawa University, Miki, Kagawa 761-07, Japan; 2, Institute of Pharmacology, 27 Taiping Road, Beijing 100850, China; 3, Institute of Endemic Bone Disease, Xian University, Xian, Shaanxi Province, China; 4, Division of Prevention and Cure for Kashin-Beck Disease, Linfen Prefecture, Shanxi Province, China.

The Effect of Roasting on the Fate of Aflatoxin B1 in Artificially Contaminated Green Coffee Beans Micco C, Miraglia M, Brera C, Desiderio C, and Masci V Mycotoxin Research 1992, Vol. 8 (2), 93-97

A study was undertaken to evaluate aflatoxin B1 contamination in coffee beans. 41 samples of green coffee were collected from large lots of material by representative sampling. The raw samples were analyzed and showed no detectable levels of aflatoxin B1. In order to establish the heat stability of the toxin, 3 artificially contaminated samples (average level l0 μg/kg) were roasted at ca 200 °C for different operation times periods so as to reproduce light and dark roasting procedures. Each sample was roasted both electrically and by gas. The percentage of toxin destruction was up to 93% for light roasted and 99% for dark roasted coffee with a slightly higher rate up to 100% for the electrically roasted coffee for light and dark roasting. In order to evaluate the potential migration of the aflatoxin B1 into the coffee beverage, 1 sample found contaminated after roasting treatment (0.8μg/kg) was extracted using each of the 3 most common types of coffee makers. Additional destruction of the toxin was observed (up to 99%) in two cases while only 75% of fate was obtained in the third. The process from raw coffee beans to beverage showed a meaningful destruction of aflatoxin B1, ranging from 97 to 100% depending on the extraction technique adopted in the preparation of the beverages. Address: Istituto Superiore di Sanità - Laboratorio Alimenti - Reparto Chimica dei Cereali, Viale Regina Elena, 299, 00161 Rome, Italy.

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Evaluation of the Extraction and Purification Procedures of the Maleyl Derivatization HPLC Technique for the Quantification of the Fumonisin B Mycotoxins in Corn Cultures Albert JF, Gelderblom WCA, and Marasas WFO Mycotoxin Research 1993, Vol. 9 (1), 2-12

The extraction and purification methods used in the maleyl derivatization HPLC technique was evaluated with respect to the pH of the extraction mixture, the extraction solvent and the purification methods used in order to determine optimum conditions for quantification of fumonisins B1, B2, and B3 in corn cultures. The highest recovery of the three compounds was obtained by extraction at pH 3.5 with CH3OHH2O (3 : 1), whilst the subsequent solvent partitioning and reversed-phase C18 Sep-pak purification have been shown to be very important in the quantification of the fumonisins in the corn cultures. The percentage recovery of the improved technique, utilizing a gradient HPLC solvent system for the simultaneous determination of the fumonisins, was 93.4 % for FB1, 68.0 % for FB2 and 82.6 % for FB3. The study indicates that the polarity of the fumonisins and consequently their solubility during extraction as well as their behavior during the subsequent purification step play an important role in quantification of these mycotoxins in corn cultures. Address: Program for Mycotoxins and Experimental Carcinogenesis, South African Medical Research Council, PO Box 19070, Tygerberg 7505, South Africa. Isolation, Structural and Toxicological Characterization of Three New Mycotoxins Produced by the Fungus Aureobasidium pullulans Schrattenholz A1 and Flesch P2 Mycotoxin Research 1993, Vol. 9 (1), 13-21

Three substances, B1, B2, and E1 were isolated from culture medium extracts of Aureobasidium pullulans by reversed phase liquid chromatography and subsequent liquid chromatographic purification steps on silica gel. The three compounds inhibited the metabolism of Saccharomyces cerevisiae and showed toxic effects in the growth inhibition test to Escherichia coli and Bacillus subtilis. Elementary analysis and mass spectroscopical methods revealed sum formulas of C23H22O6, C22H20O6, and C24H28O3 for B1, B2, and E1 and molecular weights of 394, 380, and 364, respectively. Mass spectroscopical, UV-, IR-, 13C-NMR, and 1H-NMR - spectroscopical investigations revealed polycyclic, non-aromatic compounds containing several carbonyl functions and double bonds and, most notably, spiroepoxy-functions, in the case of B1 and B2. Addresses: 1, Institut für physiologische Chemie, Universität Mainz, D-6500 Mainz, Germany; 2, Institut für Biochemie, Universität Mainz, D-6500 Mainz, Germany. Inhibition of Aflatoxin-Induced Liver Damage in Ducklings by Food Additives Soni KB1, Rajan A2, and Kuttan R1 Mycotoxin Research 1993, Vol. 9 (1), 22-26

Inhibitory effects of food additives on toxicity induced by aflatoxin B1 was conducted in 3-day-old ducklings. Aflatoxin B1 at a dose of 5 μg/day per animal for 14 days induced severe liver damage which included necrosis, fatty changes, and biliary hyperplasia. These changes were found to be inhibited by the daily administration of turmeric (50 mg), curcumin (10 mg), and ellagic acid (10 mg) in the diet. Addition of BHA - butylated hydroxy anisole (10 mg), BHT - butylated hydroxy toluene (10 mg), garlic (500mg), and asafoetida (50 mg) inhibited necrosis and degeneration of the tissue, while biliary hyperplasia persisted. Biochemical and haematological parameters were not significantly altered under the conditions studied. Addresses: 1, Arnala Cancer Research Centre, Amala Nagar Trichur, India; 2, Dept of Pathology, Veterinary College, Mannuthy, Trichur, India. 221

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A Limited Survey of Fumonisins in Corn and Corn-based Products in Asian Countries 1,2

1,2

1,2

2

2,3

4

5

6

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Ueno Y , Aoyama S , Sugiura Y , Wang DS , Lee US , Hirooka EY , Hara S , Karki T , Chen G , 7 and Yu SZ Mycotoxin Research 1993, Vol. 9 (1), 27-34

Natural occurrence of fumonisins B1 (FB1) and B2 (FB2), a promoter for hepatocarcinogenesis, was investigated in corn and corn-based products sampled in Japan, Nepal, and China by high-performance liquid chromatographic method. From the 9 imported corn kernel and 6 gluten feed samples, FB1 was detected in 8 corn (0.64.1 μg/g) and all gluten feed (0.32.4 μg/g) samples, while FB2 was found in the same corn (0.310.2 μg/g) and 3 gluten feed (0.88.5 μg/g) samples. ELISA analysis also revealed the contamination of aflatoxin B1 in 2 corn and all gluten feed samples along with fumonisins. Of 17 corn grit samples, 14 and 5 samples were contaminated with fumonisin B1 and B2 with maximum levels of 2.6 and 2.8 μg/g, respectively. As for corn-based foodstuffs marketed in Japan, no significant contamination of fumonisins was observed. Among 24 corn kernel samples in Nepal, 12 and 7 samples were positive for FB1 and FB2, and averaged to 0.6 and 1.6 μg/g, respectively. One sample showed the highest fumonisin contents as 4.6 and 5.5 μg/g, respectively. In corn samples harvested at Shanghai and Beijing, China, FB1 and FB2 were detected in various concentrations. Mycological survey has also revealed the presence of a fumonisin-producing fungus in a crop field of Japan. These findings have for the first time demonstrated high levels of contamination of fumonisins in corn and corn-based products in Asian countries. Natural co-occurrence of fumonisins and aflatoxin B1 was also detected in raw materials for mixed feed. Addresses: 1, Faculty of Pharmaceutical Sciences, Science University of Tokyo, lchigaya, ShinjukuKu, Tokyo 162, Japan; 2, Research Institute for Biosciences, Science University of Tokyo, Noda, Chiba, Japan; 3, Department of Food Engineering, National Chung-Ju Technical College, JungwonGun, Chung-Buk, Korea; 4, Centro de Ciencias Agrarias, Fundacao Universidade Estadual de Londrina, Parana, Brazil; 5, Hokkaido National Agricultural Experiment Station, Memuro-Cho, Kasai -Gun, Hokkaido, Japan; 6, Central Food Research Laboratory, Kathmandu, Nepal; 7, Institute for Public Health, Shanghai Medical University, Shanghai, China.

Effects of Naled and Dichlorvos on Growth and Production of Luteoskyrin by Penicillium islandicum Tseng HH1 and Tseng TC2 Mycotoxin Research 1993, Vol. 9 (1), 35-40

Two organophosphorus insecticides, 1,2-dibromo-2,2-dichloroethyl dimethyl phosphate (naled) and dimethyl 2,2-dichlorovinyl phosphate (dichlorvos) were used for investigation of their effects on growth and production of luteoskyrin mycotoxin by Penicilliun islandicum in various cultural media at 25 °C for 30 days or 60 days. When the concentration of naled and dichlorvos in Czapek solution broth reached 5 mg/50 mL, growth and production of luteoskyrin by the fungus was completely inhibited. In unpolished rice medium 15 mg/50 g of naled was required to retard fungal growth, while the concentration to inhibit the biosynthesis of luteoskyrin was l0 mg/50 g. On the other hand, if the medium contained dichlorvos at the level of 30 mg/50 g, the ability to produce luteoskyrin by P. islandicum was significantly reduced. In the unhulled rice case, both naled and dichlorvos at the concentration of 15 mg/50 g were necessary to retard the fungal growth, and 1 mg/50 g of each compound exhibits its ability to inhibit the toxin production. Furthermore, it was also found when the cultural medium contained only small amounts of naled and dichlorvos [0.5 mg/50 g (mL)] the capability to synthesize luteoskyrin by the fungus was drastically reduced. These data strongly suggest that both naled and dichlorvos have similar ability to inhibit luteoskyrin biosynthesis by P. islandicum and are also able to retard the fungal growth. Addresses: 1, National Laboratories of Foods and Drugs, Department of Health, Executive Yuan, Taipei, Taiwan, ROC; 2, lnstitute of Botany, Academia Sinica, Nankang, Taipei, Taiwan, ROC. 222

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Reexamination of Fusarium graminearum NRRL-13820 and NRRL-13852 Reported as Type A Trichothecene Producers Logrieco A1, Vesonder RF2, and Bottalico A3 Mycotoxin Research 1993, Vol. 9 (1), 41-46

NRRL-13820 and NRRL-13852 are reported to be two atypical Fusarium graminearum strains type A trichothecene producers [T-2 toxin (T-2) and diacetoxyscirpenol (DAS)]. These two strains were reexamined by morphological, genetical (DNA / DNA relatedness) and toxicological techniques and compared with 28 wild F. graminearum isolates obtained from corn in Italy and the USA. The isolate NRRL-13820 was morphologically confirmed as a typical isolate of F. graminearum, while the isolate NRRL-13852 showed some peculiar characteristics. Nuclear DNA comparison between NRRL-13820 and NRRL-13852 displayed 49 % similarity and showed 94 % and 44 % relatedness, respectively, when compared with F. graminearum NRRL-13833, which is a well assessed type B trichothecene producer [deoxynivalenol (DON) and 15-acetyldeoxynivalenol]. NRRL-13820, NRRL-13852, and NRRL-13833, as well as the 28 wild isolates, were not able to synthesize T-2, HT-2 nor DAS. Finally, NRRL-13820 and NRRL-13833, but not NRRL-13852, were able to produce DON (120 and 40 μg/g, respectively). The data support the concept that the production of examined type A trichothecenes is very rare in F. graminearum. Addresses: 1, Istituto tossine e micotossine da parassiti vegetali, CNR, Viale Einaudi 51, 70125 Bari, Italy; 2, Northern Regional Research- Center, ARS, 1815 University Street, Peoria, IL 61604, USA; 3, Istituto di Patologia vegetale dell‘ Università, Via E. de Nicola, 07100 Sassari, Italy. Detection and Estimation of Aflatoxins Using Both Chemical and Biological Techniques Refai MK1, Hatem ME1, Sharaby E1, and Saad MM2 Mycotoxin Research 1993, Vol. 9 (1), 47-52

Production of aflatoxins on both natural (rice and corn) and semisynthetic (VES) media was conducted using an identified toxin-producing strain of Aspergillus flavus. The A. flavus strain was able to produce 4 types of aflatoxins, namely B1, B2, G1, and G2 on rice, corn, and VES media. Quantitative data showed that the concentrations of aflatoxins B1 and G1 produced were 52, 40.3, and 39.6; and 64.7, 45.0, and 58 μg for 50 g of rice, corn, and YES media, respectively. In comparison, the yielded amounts of aflatoxins B2 and G2 were much lower: 11.5, 17.9, and 17.5; and 28.9, 40.3, and 39.5 μg for 50 g of rice, corn, and VES media, respectively. A bioassay was conducted using the following 5 standard bacterial strains: Bacillus megaterium, Bacillus subtilis, Streptococcus faecalis, Staphylococcus epidermidis, and Paracoccus denitrificans as well as a field strain of Candida albicans. All strains except P. denitrificans showed varied degrees of inhibition when applied with crude aflatoxins at 5 to 40 μg/mL. The minimum concentration of crude aflatoxins needed to inhibit P. denitrificans was l0 μg/mL. Moreover, Candida albicans was not inhibited at any concentration of aflatoxins applied in this work. Both undiluted and diluted (1/10, 1/100, 1/1000) bacterial broth cultures showed a direct relationship between the diameter of inhibition zones and the concentrations of crude aflatoxin. Mean diameters of (7.0-20.5), (5-14), (4,5-13), (3.0-12.0), and (1.5-11.0) mm were observed when various concentrations of aflatoxins were applied using B.megaterium, S.epidermidis, S.faecalis, B.subtilis, and P.denitrificans, respectively. Field trials were applied to testify the validity of our data. A 1/100 dilution was prepared from each strain of 4 different species to estimate aflatoxins in samples of contaminated corn. Both chemical and biological assays were carried out at the same time. Data revealed that the most sensitive organism inhibited by as low as 7,5μg aflatoxins/mL was B. megaterium giving an inhibition zone of 10.5mm, followed by S. epidermidis with an inhibition zone of 7.5mm. In relation, the other 2 organisms were less sensitive to crude aflatoxins. Similarly, the biological assay was applied to detect aflatoxins in some samples of wheat, corn, peanut, rice, and poultry rations. Of the 14 wheat and 10 corn samples, only 4 wheat and 2 corn samples were found to be positive. The same restults were obtained using TLC analysis. Addresses: 1, Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt; 2, Mycotoxins Lab, National Research Centre, Cairo, Egypt. 223

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Direct and Indirect Competitive Monoclonal Antibody-based ELISA of Aflatoxin B1 in Groundnut Ramakrishna N and Mehan VK Mycotoxin Research 1993, Vol. 9 (1), 53-63

Direct and indirect competitive enzyme-linked immunosorbent assays were optimized for the determination of aflatoxin B1 in groundnut utilizing a specific monoclonal antibody developed at the University of Strathclyde, UK. The monoclonal antibody was conjugated to horseradish peroxidase (HRP) for direct competitive assay, while a commercially available goat-antimouse IgG-HRP conjugate was employed for indirect competitive ELISA. Both ELISA detected aflatoxin B1 as low as 20 pg/well. Methanol-water-KCl (70 + 30 v/v, 0.5 %) extracts of groundnut were assayed by ELISA after diluting 1: 10 with PBS-Tween buffer or subjected to simple cleanup for 5 : 1 concentration prior to assay. The mean recoveries from groundnut spiked with 10 to 200 μg/kg of pure aflatoxin B1 were >90% in either ELISA, but the toxin recoveries at concentrations of 15 μg/kg were only 6567 % when subjected to cleanup and concentration before assay. The mean within-assay, inter-assay, and sub-sample coefficients of variation by ELISA of aflatoxin B1 in naturally contaminated groundnuts were, respectively, 8.9 %, 11.1 %, and 7.9 % for direct competitive assay and 4.6 %, 11.2 %, and 8 % for indirect competitive assay. Both ELISA methods are useful for routine analysis of aflatoxin B1 in groundnuts. Address: Legumes Pathology, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru PO, AP 502324, India. Moniliformin in Wheat and Triticale Grain H Lew1, J Chelkowski2, W Wakulinski2, and W Edinger1 Mycotoxin Research 1993, Vol. 9 (2), 66-71

Fusarium avenaceum infected wheat and triticale heads in Poland in each season between 1985 and 1989. The average number of heads infected by F. avenaceum was 26 % for wheat and 46 % for triticale out of all examined heads with Fusarium head blight symptoms. Fusarium-damaged wheat grain, naturally infected by F. avenaceum, contained an average of 15.9 r 7.7 mg moniliformin/kg, healthy looking kernels from the same heads an average of 0.42 r 0.19 mg moniliformin/kg. Fusariumdamaged kernels of triticale contained an average of 3.5 mg moniliormin/kg while healthy looking kernels from the same ears contained 0.25 mg/kg. Addresses: 1, Federal Institute of Agrobiology, A-4020 Linz, Austria; 2, Department of Plant Pathology, Agricultural University, P-02766 Warsaw, Poland. Occurrence of Zearalenone in Maize E Oldenburg Mycotoxin Research 1993, Vol. 9 (2), 72-78

The occurrence of zearalenone in whole plants and parts of maize usually used for silage making was investigated during the cultivation period of the crop. Zearalenone was detected up to several hundreds of μg/kg dry matter, that mainly accumulated at the end of the ripening process thus contaminating the silages subsequently. The highest concentrations of zearalenone were observed in the leaves especially at the bottom of the plant, which may correlate with leaf necrosis increasing towards harvest. Further research is needed to clarify health risks for farm animals due to uptake of contaminated feed and possible carry-over of zearalenone and its metabolites into the food chain of man. Address: Institute of Grassland and Forage Research, Federal Agricultural Research Centre, Bundesallee 50, D-38116 Braunschweig, Germany. 224

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Effect of Zearalenone on Some Physiological Processes of Maize Seeds (Zea mays L var Suwan composite) KK Sinha, G Prasad, and N Kumar Mycotoxin Research 1993, Vol. 9 (2), 79-84

Physiological processes of maize seeds (Zea mays L var Suwan composite) were found to be affected by five different concentrations (linear relationship between 100 and 2,000 μg/L) of zearalenone and duration of soaking i.e. 20 h. Inhibitions in seed germination (1458 %), shoot length (1661 %)‚ root length (976 %)‚ total chlorophyll (645 %)‚ and carotenoid (573 %) were evident due to above treatments. Address: Mycotoxin Laboratory, University Department of Botany, Bhagalpur University, Bhagalpur - 812007, India.

The Impairing Effects ot Chaetochromin D on Mitochondrial Respiration and Structure Satoru Mori1, Kiyoshi Kawai2, Yoshinori Nozawa1, Kiyotaka Koyama3, and Shinsaku Natori3 Mycotoxin Research 1993, Vol. 9 (2), 85-93

Chaetochromin D, a toxic secondary metabolite of Chaetomium gracile, was examined for impairing effects on mitochondrial respiration and structure (swelling-induction) using isolated rat liver mitochondria to gain insight into the molecular mechanism for its cytotoxicity. Chaetochromin D exerted similar mode of effects to those of chaetochromin A, cephalochromin, and ustilaginoidin A on mitochondrial reactions, causing uncoupling of oxidative phosphorylation, depression of state 3 respiration, and induction of drastic swelling in mitochondria. Chaetochromin D induced the same style of swelling as that induced by chaetochromin A, being characterized by a very high rate and small amplitude of swelling. The swelling terminated in the middle and the amplitude was about half of the full swelling. Once the quick swelling ceased in the middle, subsequent swelling could not be elicited by the second addition of chaetochromin D at any of the concentrations tested. Addresses: 1, Department of Biochemistry, Gifu University School of Medicine, Tsukasa-cho 40, Gifu 500, Japan; 2, Department of Food and Nutrition, Chukyo Women‘s University, Yokone-cho, Ohbu 474, Japan; 3, Department of Pharmacognosy, Meiji College of Pharmacy, Yato-cho, Tanashi, Tokyo 188, Japan.

Occurrence of Ochratoxin A in Herbal Drugs of Indian Origin  A Report AK Roy and S Kumar Mycotoxin Research 1993, Vol. 9 (2), 94-98

This paper contains a report of occurrence of ochratoxin A in some common herbal medicines collected from different store-houses and shop-keepers of Bihar, India. Of 129 samples of 9 plants, 55 were found to be contaminated with various levels of ochratoxin A. The level of ochratoxin A was found maximal in barks of Holarrhena antidysenterica (1.142.34 μg/g) whereas it was minimal in rhizomes of Tacca aspera (0.30.74 μg/g). Aspergillus ochraceus, A. sulphureus and Penicillium viridicatum isolates obtained from drug samples were also examined for their toxigenic potentials. 19 isolates of A. ochraceus, 13 of A. sulphureus and 37 isolates of P. viridicatum were found to be toxigenic out of 67, 33, and 107 isolates, respectively. The ochratoxin A produced by A. ochraceus was in the range of 0.09 to 2.44 μg/mL, by A. sulphureus 0.1 to 1.76 μg/mL, and by P. viridicatum 0.14 to 2.78 μg/mL of the culture filtrate. Address: Medicinal Plant Research Laboratory, University Department of Botany, Bhagalpur University, Bhagalpur - 812 007, India. 225

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Deoxynivalenol in Commercial Beer  Screening for the Toxin with an Indirect Competitive ELISA Niessen L, Böhm-Schraml M, Vogel H, and Donhauser S Mycotoxin Research 1993, Vol. 9 (2), 99-109

Antibodies specific for deoxynivalenol (DON) were prepared by immunizing rabbits with a deoxynivalenol-hemiglutaryl-HSA conjugate. The antibody showed cross reactivity with DON, 15a-acetylDON (15-Ac DON), and 3a-acetyl-DON (3-Ac DON) of 100 %‚ 216 %‚ and 260 %, respectively. No cross-reactivity was observed against 20 further trichothecenes of the A-, B-, and C-type. An indirect competitive ELISA procedure was set up for the detection of DON in liquid matrices occurring in the brewing process, including beer. ELISA was sensitive down to 3.8 ng/mL. Detection limit for DON in tenfold diluted beer was 50 ppb. Recovery of the toxin in spiked beer was 45 %, 77 %, and 83 % at toxin levels of 50; 500; and 1,000 ppb, respectively. ELISA results corresponded quite well with HPLC data. 196 commercial beers from several German breweries, including 36 gushing-positive samples, were assayed for DON. Screening revealed toxin levels from not detectable to 569 ppb. 69 % of the gushing beers had marked levels of DON. 37% of the gushing-negative beer samples contained DON in detectable amounts with a maximum content of 112 ppb. Wheat beers had significantly higher DON-contents than beers derived from barley. Concentration of the toxin was significantly higher in gushing beers than in non-gushing beers from both cereal sources. Address: Technische Universität München-Weihenstephan, Lehrstuhl für Technische Mikrobiologie und Technologie der Brauerei II, D-85350 Freising, Germany.

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Determination of Gliotoxin in Samples Associated with Cases of Intoxication in Camels M Gareisl and U Wernery2 Mycotoxin Research 1994, Vol. 10 (1), 2-8

Suspected cases of mycotoxicoses, characterized by clinical symptoms such as diarrhoea, haemorrhaging and death have been observed in breeding camels in U.A.E.. Hay samples, body fluids and intestinal contents were investigated for the presence of mycotoxins using a cell culture bioassay (MTT-test) and/or physico-chemical methods such as high performance liquid chromatography (HPLC) and mass spectrometry (MS). Extracts of the hay samples proved to be highly cytotoxic towards the swine kidney monolayers used as target cells in the bioassay. Subsequent analyses of the extracts showed the presence of the epidithiodioxopiperazine mycotoxin gliotoxin up to 0.49 mg/kg in the hay, which is the first proven case on the natural occurrence of this mycotoxin in feed. Trace amounts of ochratoxin A were also detected, while other mycotoxins known to occur naturally (i.e. aflatoxins or trichothecenes) could not be found. Gliotoxin positive HPLC-peaks were also found in some specimens (contents of rumen and intestine, allantois fluid) from camels. Gliotoxin is characterized by a variety of biological activities including antibiotic, antifungal, antiviral, cytotoxic and immunotoxic effects. Based on the results obtained, the involvement of gliotoxin as one causative agent of the intoxications is supposed. Addresses: 1, Institute for Medical Microbiology, Infectious and Epidemic Diseases, Veterinary Faculty, University of Munich, Germany; 2, Central Veterinary Research Laboratory, Dubai, U.A.E. Determination of Fumonisins in Corn: Evaluation of Two Purification Procedures Stockenström S, Sydenham EW, and Thiel PG Mycotoxin Research 1994, Vol. 10 (1), 9-14

The performance of two solid phase extraction (SPE) purification procedures, used in the determination of fumonisin B1 (FB1), B2 (FB2) and B3 (FB3) in corn, was evaluated using both thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). Fewer interferences were observed in extracts prepared using the strong anion exchange (SAX) media, in contrast to those purified on C18 media, where on occasions, visual discernment of the TLC bands was hampered by the presence of interfering compounds. Precipitate formation, resulting in the blocking of SPE cartridges was also encountered when using the C18 procedure. HPLC analyses of extracts prepared by both media indicated that they gave comparable fumonisin recoveries from naturally contaminated corn samples. The results suggest that the C18 procedure, originally developed for the TLC analyses of FB1 in mixed feeds, may also be applied to the determination of FB2 and FB3. However, where TLC is used quantitatively for fumonisin levels <1 μg/g, purification of sample extracts on SAX media is recommended. Address: Programme on Mycotoxins and Experimental Carcinogenesis, Medical Research Council, P. O. Box 19070, Tygerberg 7505, South Africa. Solubility and Stability of Sterigmatocystin in Aqueous Solutions I Septien, JL Blanco, G Suarez, and MT Cutuli Mycotoxin Research 1994, Vol. 10 (1), 15-20

In the present work we studied the ability of phosphate buffer to solubize sterigmatocystin (ST) at different pH values. We observed a higher solubility of ST at acid pH values, specially in pH 4.5. The ST adsorbed to glass was maximum at alkaline pH values. Also we studied the stability of different concentrations of ST in phosphate buffer at different pH values. After 24 h, ST can be considered stable at alkaline, neutral and 3.5 pH values. At pH 4.5, 5.5 and 6.5, ST recovered after 24 h was lower, 227

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specially at pH 4.5, when only 44 % was recovered. At day 7, ST was stable only at pH 7.5 (91 %). The lowest recovery was at pH 4.5(25 %). Address: Departamento Patología Animal I (Sanidad Animal), Facultad de Veterinaria, Universidad Complutense, 28040 Madrid, Spain. Natural Occurrence of Mycotoxins other than Aflatoxin in Africa, Asia and South America J Beardall and JD Miller Mycotoxin Research 1994, Vol. 10 (1), 21-40

This is a compilation of data on the occurrence of mycotoxins other than aflatoxin in foods and feeds from Africa, Asia and South America with 75 references. Address: Plant Research Centre, Agriculture Canada, Ottawa, Ontario K1A 0C6, Canada. Screening for Fusarium Toxigenic Isolates Using Larvae of Galleria mellonella Mulè G1‚ Logrieco A1, Bottalico A2, and Stea G1 Mycotoxin Research 1994, Vol. 10 (1), 41-46

The toxicity of twenty-four Fusarium isolates was evaluated towards larvae of Galleria mellonella by the antifeedant activity of the autoclaved fungal culture incorporated into insect diet. Cultures of isolates belonging to F. sporotrichioides, F. compactum and F. sambucinum‚ well known as capable to synthesize trichothecenes, showed high antifeedant activity (ca 6070 %); whereas controversial activity was showed by F. equiseti, F. proliferatum and F. solani cultures, and not significant activity was showed by F. oxysporum cultures. The G. mellonella larvae bioassay is suggested as a reliable sensitive insect bioassay for screening fungal toxicity. Addresses: 1, Istituto Tossine e Micotossine da parassiti vegetali, Consiglio Nazionale delle Ricerche, Bari, Italy; 2, Istituto di Patologia vegetale, Università degli Studi, Sassari, Italy.

Aspergillus flavus and Other Mycoflora of Groundnut Kernels in Israel and the Absence of Aflatoxin N Lisker1‚ R Michaeli1,2, and ZR Frank2 Mycotoxin Research 1994, Vol. 10 (1), 47-55

More than 300 groundnut (peanut) samples collected from different regions of Israel were examined by ELISA for aflatoxin contamination. Samples were designated for export, local consumption or for sowing. None of the samples were contaminated with the toxin. However, when kernels were kept at high humidity (RH  99 %), aflatoxin could be frequently detected seven days after incubation and the toxin was not uniformly distributed among kernels. Aspergillus niger, A. flavus, Penicillium citrinum and P. pinophilum were the dominant fungi and no differences were observed among cultivars. Almost half of the commercial samples examined were devoid of A. flavus. Other fungi identified were A. tamarii, A. amstelodami, P. rubrum, Rhizoctonia solani, Macrophomina phaseolina, Rhizopus spp., Sclerotium rolfsii, Fusarium and Alternaria spp.; the two last ones comprising a group of low incidence. Although groundnut samples that contain A. flavus-infected kernels are moderately common, the local climate and agrotechniques in use in Israel are not conducive to aflatoxin accumulation. Nevertheless infected kernels may become a threat to health if stored under inadequate conditions. Addresses: 1, Department of Agronomy and Natural Resources, The Volcani Center, Agricultural Research Organization, Bet Dagan 50250, Israel; 2, Department of Plant Pathology, The Volcani Center, Agricultural Research Organization, Bet Dagan 50250, Israel. 228

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Mycotoxin Production by Some Indian Alternaria Species KS Bilgrami, AA Ansari, AK Sinha, AK Shrivastava, and KK Sinha Mycotoxin Research 1994, Vol. 10 (1), 56-59

A total of seven species of Alternaria: A. alternata (Fr.) Keissler; A. capsici-annui Savul & Sandu; A. citri Ellis & Pierce; A. porri (Ellis) Cifferi; A. radicina Meier, Drechsler & Eddy; A. tenuissima (Kunze : Pers) Wiltshire and A. tomato (Cooke) Jones were screened on rice culture medium for their ability to elaborate five major Alternaria mycotoxins viz. tenuazonic acid (TA), alternariol (AOH), alternariol methyl ether (AME), altenuene (ALT) and altertoxin-I (ATX-I). All the species produced mycotoxins in varying concentrations. A. capsici-annui was recorded as the mycotoxin producer for the first time. ALT by A. citri and A. tomato; ALT, and ATX-I by A. tenuissima; ALT, TA and AME by A. porri and TA by A. radicina are the new additions to the list of mycotoxins produced by the respective species of Alternaria. Address: University Department of Botany, Bhagalpur University, Bhagalpur-812 007, India.

Effects of Mycotoxins on the Mixed Lymphocyte Reaction Itoh T, Ishii K, and Ueno Y Mycotoxin Research 1994, Vol. 10 (2), 62-66

Effects of toxic fungal metabolites on mixed lymphocyte reaction (MLR) using mouse splenocytes and on growth of mouse myeloma cells were examined. Among 25 toxins assayed, the IC50 values of emodin, luteoskyrin, sterigmatocystin, deoxynivalenol, 4-acetylnivalenol, T-2 toxin and fusaric acid for the MLR were lower than those for the cytotoxicity toward the myeloma cells, suggesting that these toxins possess suppressive activity to the cellular immune system. Address: Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Shinjuku-ku, Tokyo 162, Japan.

Production of Fumonisin B1 and B2 by Fusarium moniliforme Isolated from Korean Corn Kernels for Feed Lee Ung-Soo1, Lee Myong-Yur2, Shin Kwang-Sop3, Min Yun-Sik1, Cho Chae-Min1, and Yoshio Ueno4 Mycotoxin Research 1994, Vol. 10 (2), 67-72

The natural occurrence of fumonisin B1 (FB1) and B2 (FB2), a promoter for hepatocarcinogenesis, was investigated in Korean corn kernels for feed by HPLC with fluorescence detection. From the 12 corn kernel samples, FB1 was detected in 5 samples at levels ranging from 53 to 1327ng/g, while FB2 was found in 4 samples from 69 to 680ng/g. In the positive samples, the average concentrations of FB1 and FB2 were 506 and 288ng/g, respectively. One sample (No. K3) showed the highest FB1 and FB2 contents as 1321 and 680ng/g, respectively. In the micrological survey on 5 positive samples for FB1 and FB2, 6 strains of Fusarium moniliforme were isolated, and all these isolates had a producibility of FB1 and FB2, with maximum levels of 80.7 to 180.9 g/g. This is the first report on the natural cocontamination of FB1 and FB2 in Korean corn kernels for feed, and on the ability of F. moniliforme isolated from corn kernels for feed in Korea to produce FB1 and FB2. Addresses: 1, Department of Food Engineering, National Chung-Ju University, Jungwon Gun, ChungBuk 383-870, Korea; 2, Department of Food Engineering, Chung-Buk National University, Cheong Ju, Chung-Buk 360-763, Korea; 3, Department of Mechanical Engineering, National Chung-Ju University, Jungwon Gun, Chung-Buk 383-870, Korea; 4, Department of Toxicoiogy and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Ichigaya, Shinjuku-ku, Tokyo 162 Japan. 229

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Production of Beauvericin by Fusarium proliferatum from Maize in Italy 1

1

2

3

3

A. Moretti , A. Logrieco , A. Bottalico , A. Ritieni , and G. Randazzo Mycotoxin Research 1994, Vol. 10 (2), 73-78

Beauvericin (BEA), a toxic cyclodepsipeptide, was purified from corn kernel cultures of a toxigenic strain of Fusarium proliferatum, isolated from corn ear rot in northern Italy (45 mg/kg dry culture). The strain, designed ITEM-1503, also produced fumonisin B1 (2,250 mg/kg dry culture), and moniliformin (150 mg/kg dry culture). Thin-layer chromatography, high-performance thin layer chromatography, high performance liquid chromatography, low-resolution electronic impact mass spectrometry and 1H, 13C nuclear magnetic resonance spectroscopy were used for BEA isolation and confirmation. This is the first report on the production of BEA by F. proliferatum. Addresses: 1, Istituto Tossine e Micotossine da parassiti vegetali, Consiglio Nazionale delle Ricerche, 70125 Bari; 2, Istituto di Patologia vegetale, Universitá degli Studi, 07100 Sassari; 3, Dipartimento di Scienze degli alimenti, Universitá degli Studi di Napoli “Frederico II“, 80055 Portici, Italy.

Production of Alternariol and Alternariol Monomethyl Ether in Natural Substrates in Comparison with Semisynthetic Culture Medium S Chulze, A Torres, A Dalcero, and M Combina Mycotoxin Research 1994, Vol. 10 (2), 79-84

The comparison in toxins production and growth by Alternaria strains in liquid, solid culture media and natural substrates (rice and sunflower) was evaluated. Ground rice- corn steep liquor medium (GRCS) was the more suitable medium for production of alternariol (AOH) and alternariol monomethyl ether (AME). The maximum levels produced were 676 μg/50 ml AOH and 1570/50 ml AME. Rice was better than sunflower in supporting toxins production. Different ratios AOH/AME were found according to the substrate evaluated. Address: Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico- Químicas y Naturales, Universidad Nacional de Río Cuarto, Córdoba, Argentina.

Subclinic Effect of the Administration of T-2 Toxin and Nivalenol in Mice Pacin A1,2, Reale C3‚ Mirengui H3‚ Orellana G4, and Boente G4,5 Mycotoxin Research 1994, Vol. 10 (2), 85-96

Four experiments using T-2 toxin and nivalenol at different dosage, which represented the 25% and 40% of the LD50 (experiment A: 1.04 mg of T-2 toxin per kilogram of body weight, experiment B: 2.34 mg of T-2 toxin/kg b.w., experiment C: 1.04 mg of T-2 toxin/kg b. w. and 2.34 mg of T-2 toxin/kg b.w.; experiment D: 0.82 mg of nivalenol/kg b.w. and 1.845 mg of nivalenol/kg b.w.) were conducted on 400 mice. Both toxins were administered to mice of different ages (experiments A and B were adults, experiment C and D were young) by intraperitoneal single injection, and the clinical signs, hematological variables and histoanatomo-pathological changes were studied. All animals survived. No changes anatomo-histopathological nor significative differences in weight gain were observed. Different behaviors were found for nivalenol and T-2 toxin. The most significant change was the increase in the level of monocytes in old animals, so this could be a biological indicator for T-2 toxin subclinical intoxication. Addresses: 1, Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CIC); 2, Universidad Nacional de Luján; 3, Universidad Nacional del Sur; 4, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (UBA); 5, Consejo Nacional de Investigaciones Cientificas y Técnicas (CONICET).

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Aflatoxins B1 and G1 Solubility in Standard Solutions and Stability During Cold Storage ME Garcia, JL Blanco, and G Suarez Mycotoxin Research 1994, Vol. 10 (2), 97-100

In the present work we study the use of different solvents to store aflatoxins B1 and G1 standard solutions. We have obtained significant differences between aflatoxin B1 and G1 in ethyl acetate, methanol and water, with aflatoxin G1 being less stable. We recommend chloroform as the election solvent to store the aflatoxin solutions. The fact that aflatoxins are highly stable in water may have a potential use in experiments of biological activity. Address: Departamento Patología Animal I (Sanidad Animal), Facultad deVeterinaria, Universidad Complutense. 28040 Madrid, Spain.

Production of Aflatoxin and Cyclopiazonic Acid by Various Aspergilli: An ELISA Analysis 1 2 1 X Huang ‚ JW Dorner and Fun S Chu

Mycotoxin Research 1994, Vol. 10 (2), 101-106

An enzyme-linked immunosorbent assay (ELISA) was used to monitor a total of 153 fungi in the Aspergillus flavus group, including 130 A. flavus, 15 A. parasiticus and 8 A. tamarii, for their ability to produce aflatoxins (AFs)and cyclopiazonic acid (CPA) in a mycological broth-sucrose-yeast extract medium. Of 15 A. parasiticus isolates, ten produced AFs in a range of 12.4 to 89.3 μg/vial (average 56.9 μg/vial); two isolates produced only trace amounts of AFs and three isolates produced none at all. Production of CPA was not demonstrated in any A. parasiticus isolate. On the other hand, all A. tamaii isolates produced only CPA with a range of 310 to 1100 μg/vial. Fifteen percent (14.6%) of the A. flavus isolates (19/130) produced more than 500 μg CPA/vial, but yielded no or little AF (less than 0.1 μg/ vial). About 22.3% of A. flavus (29/1 30) that produced less than 500 μg of CPA also yielded little or no aflatoxin. Most A. flavus isolates (44.6%) produced both CPA (50 to 300 μg/vial) and AFs (10 to 40 μg/vial). About 9.2% of the A. flavus are low CPA producers (less than 100 μg/vial) but yielded higher amounts of AFs. A small percentage (12/130 or 9.2%) of A. flavus isolates produced neither CPA nor aflatoxin. Excluding the isolates that produced neither AFs nor CPA, there is a negative correlation between the production of CPA and AFs by most A. flavus isolates. Data obtained from ELISA for the production of CPA were consistent with TLC results. Thus, the ELISA method for CPA and AFB could be applied to the screening of toxigenic fungi. Data on the simultaneous production of both toxins by a large percentage of the toxigenic A. flavus isolates suggest that there is a potential heath hazard for co-existence of both toxins in foods and feeds. Addresses: 1, Food Research Institute and Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, WI 53706; 2, National Peanut Laboratory, USDA, ARS, USDA, Dawson, GA31742.

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Evalution of Liquid Media for Fumonisin Production by Fusarium moniliforme MCR 826 JF Alberts, WCA Gelderblom, WFO Marasas, and JP Rheeder Mycotoxin Research 1994, Vol. 10 (2), 107-115

Production of fumonisins B1 (FB1) and B2 (FB2) by 5 lyophilization batches of Fusarium moniliforme strain MRC 826 was studied in several liquid media and vermiculite supplemented with liquid media. In addition the effect of different parameters including pH, inert material, shake versus stationary cultures as well as different carbon sources on the production of the fumonisins were investigated. Fumonisin production in liquid cultures was significantly (P<0.01) correlated (r = 0.920.98) with fungal growth, which in turn is affected by the pH of the medium as well as the carbon source utilized. The highest FB1 yields (approximately 40 mg/l) over the incubation period of 14 days were produced in a chemically defined medium with glucose as carbon source set at an initial pH value of 4. FB1 production in “corn patty“ cultures (approximately 1 to 3 g/kg), however, by far exceeded that obtained in the liquid media, while poor fungal growth and fumonisin production was obtained in vermiculite supplemented cultures. From these studies it became clear that the ability of a culture to produce fumonisins is determined by the interaction of a variety of physiological and nutritional factors regarding the inoculum and the culture medium. Address: Programme on Mycotoxins and Experimental Carcinogenesis (PROMEC), South African Medical Research Council, P.O. Box 19070, Tygerberg, 7505, South Africa.

Fusarium Poae (Peck) Wollenw. - Occurrence in Maize Ears, Nivalenol Production and Mycotoxin Accumulation in Cobs J Chelkowski1, H Lew2, and H Pettersson3 Mycotoxin Research 1994, Vol. 10 (2), 116-120

Fusarium poae(Peck) Wollenw. occurred in maize ears with “pink rot“ during 19851993 up to 18% of Fusarium isolates, with maximum frequency in 1990. Nivalenol and fusarenone X were produced under laboratory conditions by 13 out of 14 isolates up to 115 μg/g and 13.3 μg/g respectively. The same isolates produced diacetoxyscirpenol (DAS) up to 21.7 μg/g and 15-monoacetoxyscirpenol (MAS) up to 12.3 μg/g. However, in none of the strain cultures were T-2 toxin and HT-2 toxin detected. In samples of naturally infected maize grain nivalenol was detected at levels of 1.832.5 μg/g and fusarenone X was not present in corresponding axial stems were present both nivalenol (up to 13.5 μg/g) and fusarenone X (up to 2.4 mg/g). Addresses: 1, Institute of Plant Genetics, Polish Academy of Sciences, 60-479 Poznan, Poland; 2, Federal Institute of Agrobiology, Linz, Austria; 3, Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden.

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Aflatoxin Induced Hepatocarcinogenesis in Ducks T Leenadevi, KV Valsala, and A Rajan Mycotoxin Research 1995, Vol. 11 (1), 2-8

To assess the carcinogenic response of duck hepatic tissue, an experimental study was undertaken. Pure aflatoxin B1 was administered to twelve white pekin ducks of two to three months of age at a dose rate of 0.04165 mg/kg body weight every third day for six months. There was reduction in body weight and haemoglobin level from the third month onwards. Total serum protein, albumin and globulin had a slow and gradual reduction and ESR was significantly increased from the third month. Serum enzymes (AST, ALT, GGT and ALP) were significantly increased from the VIIIth to XVth fortnights (Two weeks). Ten ducks developed hepatic tumours by 180th day. Four of them had neoplastic nodules on the 90th day. Histopathologically they were hepatocellular carcinoma (6), Cholangiocellular carcinoma (4) and Chronic hepatitis (2). There was moderate to severe expression of GGT and ALP in the liver tissue during neoplastic transformation. Address: Centre of Excellence in Pathology, College of Veterinary and Animal Sciences, Kerala Agricultural University – Thrissur. Occurrence of Moniliformin, Deoxynivalenol, and Zearalenone in Durum Wheat (Triticum durum Desf.) A Adler1, H Lew1, W Brodacz1, W Edinger1, and M Oberforster2 Mycotoxin Research 1995, Vol. 11 (1), 9-15

Forty-eight durum wheat samples from 5 locations in Austria were examined for Fusarium infection and Fusarium toxin content. F. graminearum and F. avenaceum were by far the prevailing Fusarium species in durum wheat kernels, followed by F. poae, F. culmorum, and F. equiseti. Ion-paired HPLC analyses of the samples showed moniliformin contents of kernels up to 0.88 mg/kg. All moniliformin contaminated samples also contained high levels of deoxynivalenol (up to 8.2 mg/kg) and lower levels of zearalenone (< 0.33 mg/kg). The levels of zearalenone in naturally contaminated durum wheat samples did not correspond to the high yields of zearalenone found in cultures of the fusaria isolated from the durum wheat kernels. These conflicting results as well as some toxicological aspects of the carry over of Fusarium toxins from durum wheat kernels into pasta are discussed. Addresses: 1, Federal Institute of Agrobiology, 4020 Linz, Austria; 2, Federal Institute of Plant Production, 1020 Vienna, Austria.

Ochratoxin-A Production in Brazilian Dry Beans (Phaseolus vulgaris L.) MSC Cordeiro1, J Amaya-Farfan2, and PJS Moran3 Mycotoxin Research 1995, Vol. 11 (1), 16-20

Ochratoxin A was produced at concentrations of about 200 mg kg-1 of dry beans (Phaseolus vulgaris L.) of each of five Brazilian commercial varieties. Both intact and decorticated kernels of the varieties Preto, Branco, Rosinha, Roxo and Carioca (22% moisture) were inoculated with Aspergillus alutaceous and incubated at 25°C for 28 days. Results from thin-layer and column chromatography, mass, infrared, 1H-nuclear magnetic resonance and UV-spectrometry showed that 1) the common bean is a highly stimulatory substrate for the bioproduction of ochratoxin A and 2) the putative toxin extracted by the method of Soares & Rodriguez-Amaya was in fact ochratoxin A. Removal of the seed coat resulted in increased OTA production for all varieties, particularly for the Rosinha, Roxo and Carioca. Addresses: 1, Depto. de Química, Universidade Federal do Amazonas, Manaus Brasil; 2, DEPAN, Fac. Engenharia de Alimentos, Universidade Estadual de Campinas - UNICAMP, CP 6121, 13081970 Campinas, SP, Brasil; 3, Instituto de Química, UNICAMP, CP 6154, Campinas, SP, Brasil. 233

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Contamination of Red Chilli with Aflatoxin B1 in Pakistan Zuzzer A Shamsuddin, Mobeen A Khan, Butool A Khan, Mansoor A Ahmad, and Aftab Ahmed Mycotoxin Research 1995, Vol. 11 (1), 21-24

A survey of red chilli (Capsicum indicum) for contamination with aflatoxins was performed on different samples comprising whole, crushed and powdered red chilli collected from various stores located in the city of Karachi, Pakistan. Red chilli required rather rigorous clean-up procedure for removal of adulterants and interference resulting from various types of compounds. A modified Romer method followed by bi-directional thin layer chromatography (TLC) was used for the detection of aflatoxins and confirmatory tests were performed by spraying the TLC plates with 50% sulphuric acid and making the derivative with trifluoro-acetic acid. Of all the 176 samples of red chilli examined, 66% were found to be contaminated with aflatoxin B1. Generally, samples of red chilli exammined were found to be fairly low in aflatoxin B1 content, whereas only seven samples were found to contain concentrations greater than 25 μg/kg of aflatoxin B1. Address: Mycotoxins Laboratory, P.C.S.I.R. Laboratories Complex, Karachi-75280, Pakistan.

Identification of Nephrotoxic Penicillium Species from Cereal Grains JT Mills1, JC Frisvad2, KA Seifert3, and D Abramson1 Mycotoxin Research 1995, Vol. 11 (1), 25-35

A system is described for identifying grain-inhabiting nephrotoxic Penicillium spp. based on their colony characters on Czapek yeast extract agar, yeast extract sucrose agar, and malt extract agar media, and their secondary metabolite profiles on thin layer chromatography plates. Using this system, the identity of 11 Penicillium species, or their chemotypes, producing nephrotoxic metabolites could be confirmed. The species are P. verrucosum chemotype I, P. verrucosum chemotype II, P. expansum, P. citrinum, P. aurantiogriseum, P. freii, P. tricolor, P. polonicum, P. viridicatum, P. cyclopium, and P. melanoconidium. Other non-nephrotoxic Penicillium species present on stored grains were separated from nephrotoxic species by their colony characters and metabolite profiles. Addresses: 1, Agriculture and Agri-Food Canada, Research Centre, 195 Dafoe Road, Winnipeg, Manitoba, Canada R3T 2M9; 2, Department of Biotechnology, Technical University of Denmark, DK-2800 Lyngby, Denmark; 3, Centre for Land and Biological Resources Research, Research Branch, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6.

Detoxification of Ochratoxin A, a Food Contaminant: Prevention of Growth of Aspergillus ochraceus and Its Production of Ochratoxin A P Deberghes1, AM Betbeder1, F Boisard2, R Blanc1, JF Delaby2, S Krivobok3, R Steiman3, F Seigle-Murandi3 and EE Creppy1 Mycotoxin Research 1995, Vol. 11 (1), 37-47

The toxicity of ochratoxin A (OTA), a mycotoxin produced by fungi of Aspergillus or Penicillium genera is now well documented. Its nephrotoxicity, immunosuppression, teratogenicity, and carcinogenicity have been widely studied. Physical and biochemical methods have been studied to prevent these toxinogenic Aspergillus and Penicillium from producing OTA, and/or to destroy the mycotoxin when already produced in a liquid or a solid medium. Repeated freezing at – 20 °C and thawing at + 26 °C aleatory reduce OTA production in a liquid medium. Exposure to UV B for different periods of time is efficient in preventing OTA production in a liquid medium. Gamma-irradiation from 2 to 5 kGy gives good results in preventing the production of OTA or destroying it when already produced. Carboxypeptidase is very efficient at 5 units/50 ml in a liquid medium for cleaving the OTA already produced. 234

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Addresses: 1, Laboratoire de Toxicologie et Hygiène Appliquée, UFR de Pharmacie, 3 ter place de la Victoire 33000 Bordeaux; 2, Centre d‘Etudes Scientifiques et Techniques de l’Armée [C.E.S.T.A.], 33114 Le Barp, France; 3, Université Joseph Fourier, UFR de Pharmacie, Laboratoire de Mycologie BP 138, 38243 MEYLAN Cedex.

Influence of Water Activity on the Production of T-2 Toxin by Fusarium sporotrichioides M Schwabe and J Krämer Mycotoxin Research 1995, Vol. 11 (1), 48-52

The production of T-2 Toxin by two strains of Fusarium sporotrichioides at high (0.995), medium (0.970) and low (0.945) water activity (aw) was investigated. The organisms were incubated in an optimal liquid base medium (Wort broth) adjusted to the different aw values. With decreasing aw the amount of T-2 Toxin in the culture supernatants fell drastically. Comparing cultures that had reached almost equal mycelial dry weights, those of low aw had T-2 Toxin levels 10 to 100 times lower than those of high aw. This effect was observed using both glycerol or sodium chloride as humectants. Address: Department of Agricultural and Food Microbiology, University of Bonn, Meckenheimer Allee 168, 53115 Bonn.

Fusarium sp. FN-2B: a Controversial Strain Genetically Close to Fusarium poae 1 1 2 G Mule , A Logrieco , and A Bottalico

Mycotoxin Research 1995, Vol. 11 (1), 53-58

Fusarium strain Fn-2B, a trichothecene producing Fusarium strain, first reported as F. nivale but with a very controversial identification, was reexamined genetically by nucleotide sequencing from a highly variable region of the large subunit (25- 28S) rRNA (D2 region, ca. 220 nucleotides), and compared to the same region from species it was presumed to belong, in order to assess its phylogenetic affinity. Fusarium strain Fn-2B proved to be more closely related to F. poae NRRL-13637 showing only one heteromorphic site. In comparison to other fungal strains, Fn 2B showed 3, 11, and 34 bases that differ from F. sporotrichioides NRRL-3299, F. tricinctum NRRL-13636 and Microdochium nivale NRRL13934, respectively. This phylogenetic affinity between Fusarium strain Fn-2B and F. poae is well correlates with the production of trichothecene mycotoxins by the species. Addresses: 1, Istituto Tossine e Micotossine da Parassiti Vegetali, Consiglio Nazionale delle Ricerche, 70126 Bari, Italy; 2, Istituto di Patologia vegetale, Universitá degli Studi, Sassari 07100, Italy.

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Influence of Inoculum Size on Aflatoxin Production in Home-Made Yoghurt ME Garcia, JL Blanco, and G Suarez Mycotoxin Research 1995, Vol. 11 (2), 62-68

In previous works we have studied the influence of different factors on the aflatoxin production in yoghurt. In the present paper we complete our investigations with the study of the influence of the inoculum size. The inoculum sizes used by us were from 4 x 101 to 4 x 106. As can be expected, the fungal growth, expressed as dry mycelium weight, was lower in 4 x 101 and higher in 4 x 106. The amount of aflatoxin in the mycelium was stable, or increased slightly with the inoculum size. In the substrate, the amount of aflatoxin was stable with little fluctuations, with a higher level of toxin in 4 x 103 and lower one in 4 x 106. We detected a higher aflatoxin level in the mycelium than in the substrate. Address: Departamento Patología Animal I (Sanidad Animal), Facultad de Veterinaria, Universidad Complutense. 28040 Madrid, Spain.

Effect of Pesticides in Alternaria alternata Mycotoxins Production A Dalcero1, S Chulze1, A Torres1, M Rodriguez2, and M Combina1 Mycotoxin Research 1995, Vol. 11 (2), 69-74

The effect of pesticides on Alternaria mycotoxins was evaluated both in culture media and in sunflower seeds. Different behaviour was observed depending on whether insecticides or fungicides were considered. When Captan, Lindaphor and Dichlorvos were evaluated in sunflower seeds their effects were dependent on the toxin being evaluated (alternariol, alternariol monomethyl ether and tenuazonic acid). A full spectrum of effects was observed, ranging from no effect, stimulation, to inhibition. Addresses: 1, Departamento de Microbiologia e lnmunologia and 2, Departamento de Matematica, Facultad de Ciencias Exactas, Fisico- Quimicas y Naturales. Universidad Nacional de Rio Cuarto, Córdoba, Argentina.

Aflatoxin Production on the Pits (Seeds) of the Date Palm (Phoenix dactylifera L.) Ahmed IA1,Ahmed AK1, and Robinson RK2 Mycotoxin Research 1995, Vol.11 (2), 75-84

Pits - a by-product of the utilization of date fruits, are widely used as components of animal feeds, but an incident of aflatoxicosis in camels fed rations containing date pits has caused concern in the Gulf Region. This present study has shown that date pits can support aflatoxin production when inoculated with Aspergillus parasiticus (IMI 91019b) and that variety and/or stages of maturation within a given variety can affect the final level of aflatoxin in the material. In one variety, Lulu, aflatoxin production was 44.5, 38.7 and 21.0 μg /g in pits taken from the first three stages of ripening namely Kimri, Khalal and Rutab, but no significant aflatoxin production was noted at the fully-ripe Tamr stage. Moisture content was considered to be the most important factor with respect to the capacity of the mould to synthesise aflatoxin in date pits. Addresses: 1, Central Food Control and Consultancy Laboratory, Sharjah, United Arab Emirates; 2, Department of Food Science and Technology, University of Reading, United Kingdom.

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Trichothecene Chemotypes of Fusarium graminearum Isolated from Corn in Hungary 1

2

Szécsi Á and Bartók T

Mycotoxin Research 1995, Vol. 11 (2), 85-92

Twenty three strains of Fusarium graminearum isolated from corn were screened for their ability to produce type A and B trichothecenes and zearalenone (ZEA) on the solid substrate rice grains in the dark at 28°C for 21 days. Toxin analyses were made with HPLC technique. Of 23 total isolates, 10 produced deoxynivalenol (DON), 4 produced DON and nivalenol (NIV), 1 produced DON and 15 acetyl-DON (15-ADON), 1 produced NIV and 4-acetyl-NIV (4-ANIV) and 1 produced NIV. Of 23 total F. graminearum isolates, 20 produced ZEA. These results suggest that strains of F. graminearum, prevailing in Hungarian corn growing regions, might belong to DON- and NIV-chemotypes. This is the first report demonstrating that DON-, DON-NIV-, DON-15-ADON-, NIV-4-ANIV and NIV-producing F. graminearum isolates are distributed in Hungary. Addresses: 1, Department of Plant Pathology, Plant Protection Institute, Hung. Acad. Sci., H-1525 Budapest, P.O. Box 102, Hungary; 2, Analytical Laboratory, Cereal Research Institute, H-6701 Szeged, P.O. Box 391, Hungary.

Fusarium Species Associated with Banana Fruit Rot and Their Potential Toxigenicity RF Vesonder1, A Logrieco2, A Bottalico2, C Altomare2, and SW Peterson3 Mycotoxin Research 1995, Vol. 11 (2), 93-98

Banana fruits exhibiting signs of decay were collected from markets in the United States and Italy. Fungi isolated from the lesions on the banana fruits were Fusarium moniliforme, F. subglutinans, and F. semitectum var. majus. When the fungal strains were cultivated on maize kernels, the cultures did not produce zearalenone (ZON), zearalenols (á-, â -ZOH), and trichothecenes [deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), T-2 toxin (T-2), diacetoxyscirpenol (DAS)]. Fumonisins and fusarin C (FUS-C) were not detected naturally nor in bananas purchased in the U.S. and artificially infected with Fusarium. Moniliformin (M) (up to 267 mg/kg) was detected in maize kernel cultures of F. subglutinans from bananas. No mycotoxins were detected in naturally infected fruits. Although no mycotoxins were detected in the extracts from corn cultures of F. semitectum var. majus, the extracts were toxic to brine shrimp and mice. Addresses: 1, Mycotoxin Research, National Center for Agricultural Utilization Research, Peoria, IL 61604 USA; 2, Instituto tossine e micotossine da parassiti vegetali, Consiglio Nazionale delle Ricerche, 70126 Bari, Italy; 3, Microbial Properties Research, National Center for Agricultural Utilization Research, Peoria, IL 61604 USA.

Synthesis, Structure and Anti-Acetylcholinesterase Activity of 12a-O-Methyl Territrem B Peng Fu Chuo and Chen Ping Ho Mycotoxin Research 1995, Vol. 11 (2), 99-102

12a-O-methyl territrem B was synthesized from TRB by treatment with dimethyl sulfate in methanolic NaOH. The structure of 12a-O- methyl territrem B was elucidated by uv, ir nmr and mass spectra and it is 2.5 times more potent than TAB in inhibitory activity on electric eel acetylcholinesterase (E. C. 3.1.1.7.) (AChE). Address: Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan ROC.

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Preharvest Aflatoxin Contamination of Groundnuts Subjected to Terminal Drought Stress in Postrainy Season VK Mehan, N Ramakrishna, RCN Rao, and D McDonald Mycotoxin Research 1995, Vol. 11 (2), 103-109

Groundnuts grown in the postrainy season under terminal drought stress imposed by withholding irrigation, or under a water-deficit gradient created by line-source sprinkler irrigation, were examined for preharvest aflatoxin contamination. High levels of aflatoxin B1 were found in damaged seeds in both situations. When grown under continuous drought-stress, toxin levels in damaged seed samples ranged from 1480 to 2467 μg/kg in the 1990/91, and 1.3 to 2000 μg/kg In the 1991/92 postrainy seasons. Aflatoxin B1 contamination in all damaged seed samples increased with increasing water deficit; toxin levels ranged from 26 to 850 μg/kg across the water deficit gradient. Aflatoxin was either absent or almost negligible (1-2 μg/kg) in apparently undamaged seed samples. Low risk of aflatoxin contamination in apparently undamaged seeds of groundnuts grown in postrainy seasons is indicated, even when there is terminal drought stress. Address: Legumes Program, International Crops Research Institute for the Semi-Arid Tropics, Patancheru PO, AP 502 324, India.

Determination of Ochratoxin A by Immunoaffinity Column Clean-Up and HPLC in Wheat and Pig Liver Marley EC, Nicol WC, and Candlish AAG Mycotoxin Research 1995, Vol. 11 (2), 111-116

Determination of ochratoxin A (OTA) by immunoaffinity column clean-up and HPLC detection was performed on wheat and pig liver. Several extraction protocols involving methanol and ethyl acetate were investigated. The optimum experimental conditions for analysis of OTA in artificially contaminated wheat (87.4% recovery) using immunoaffinity column clean-up was found to be the methanol:PBS (1:1 v/v) protocol. These conditions, however, gave low recoveries for pig liver (40.4%). Address: Rhône Poulenc Diagnostics West of Scotland Science Park 3.06 Kelvin Campus, Maryhill Road Glasgow G20 0SP Scotland, United Kingdom.

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Moniliformin Production by Fusarium Species Isolated from Argentinian Corn 1

2,3

1,4

CEP Sanhueza , HHL González , and SL Resnik Mycotoxin Research 1996, Vol. 12 (1), 2-6

Forty-one isolates of Fusarium obtained from the main Argentinian corn production area were tested for their ability to produce moniliformin. One of 22 isolates of F. moniliforme, 2/10 of F. proliferatum and 3/9 of F. subglutinans, produced moniliformin in a range between 0.3 to 2.7 mg/g. These data represent the first report of the production of moniliformin by Fusarium species from section Liseola in Argentina. Addresses: 1, Departamento de Química Orgánica. F.C.E. y N.-U.B.A., Buenos Aires, Argentina; 2, Departamento de Ingeniería Química. F.I.-U.B.A., Buenos Aires, Argentina; 3, Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina; 4, Comisión de Investigaciones Científicas de la Provincia de Buenos Aires, Argentina. Production of 14C-Ochratoxin A by Penicillium verrucosum sp. 1761 in Liquid Culture M Ruhland, G Engelhardt, and PR Wallnöfer Mycotoxin Research 1996, Vol. 12 (1), 7-13

The synthesis of 14C-ochratoxin A was carried out by biosynthesis with Penicillium verrucosum sp. 1761 in liquid culture using 2-14C-Na-acetate and 2-14C-malonic acid as precursor. The largest amount of ochratoxin A produced by Penicillium verrucosum was detected in a minimal medium after an incubation period of 28 days. By addition of citric acid and 2,4-dichlorophenoxy acetic acid larger ochratoxin A contents could be measured. For transformation studies of ochratoxin A, a molecule which is exclusively labelled in the isocoumarin moiety is best suitable. This labelling could be obtained by using labelled malonic acid as precursor, whereas with 2-14C-NA-acetate as precursor the whole molecule was labelled. Incorporation rates of radioactivity into the molecule were from 0.04 % to 0.11 %‚ specific radioactivities from 0.02 μCi/mg to 0.07 μCi/mg. Address: Bayerische Landesanstalt für Ernährung, Menzingerstr. 54, 80638 Munich, Germany

Fungi and Mycotoxins in South African Maize of the 1993 Crop E Rava, JH Viljoen, H Kallmeyer, and A de Jager Mycotoxin Research 1996, Vol. 12 (1), 15-24

The most dominant fungi in South African maize for the 1993 crop were Fusarium moniliforme and Fusarium subglutinans. The mycotoxins occurring with the most frequency were the fumonisins, deoxynivalenol and nivalenol. Levels of fungal infection and mycotoxins varied between white maize and yellow maize, and between the respective grades. Address: Maize Board, P.O. Box 669, Pretoria, 0001, South Africa.

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Mycotoxins in Maize products of the 1994/95 marketing season E Rava Mycotoxin Research 1996, Vol. 12 (1), 25-30

The levels of mycotoxins found were generally lower the more refined the maize products were. The highest levels of deoxynivalenol, nivalenol and fumonisins were found in maize bran and maize screenings. Ochratoxin A was found in a defatted germ meal sample at 50 μg/kg. Zearalenone was found randomly in samples at low levels (50–100 μg/kg). All other mycotoxins tested for, except in maize bran and maize screenings, were absent above the detection limits. Address: Maize Board, P.O. Box 669, Pretoria, 0001, South Africa.

Incidence of Fusaria and Occurrence of Selected Fusarium Mycotoxins on Lolium spp. in Germany R Engels and J Krämer Mycotoxin Research 1996, Vol. 12 (1), 31-40

Test plantings with varieties of Lolium multiflorum and L. perenne were harvested 4 to 7 times a year in 1991 and 1992. Samples were checked tor the presence of Fusaria, the mycotoxins zearalenone, T2 toxin, and diacetoxyscirpenol (DAS). Spectrum of species and the incidence of Fusaria and fusariotoxins are discussed in relation to the influencing factors site, variety of Lolium, harvesting time and year. Depending on these factors, 41 % to 100% of the samples were Fusarium positive. Differences in infestation with Fusarium among varieties of Lolium perenne were dependent on location and did not correlate with yield. The six species of Fusarium pathogenic to Lolium spp. (F. graminearum, F. culmorum, F. avenaceum, F. oxysporum, F. solani, and F. acuminatum) totaled 35.7 % of all the isolated strains. 14 species could be isolated from Lolium samples (descending frequency): F. culmorum, F. sambucinum, F. equiseti, F. acuminatum, F. semitectum, F. oxysporum, F. subglutinans, F. avenaceum, F. sporotrichioides, F. proliferatum, F. tricinctum, F. anthophilum, F. dimerum and F. graminearum. For the detection of Fusaria a promising new immunological method is presented. lt is based on the genus specific production of exopolysaccharides by Fusarium species. Mycotoxin contents in grass ranged from 0.01 to 4.75 ppm for zearalenone with 67% positive samples and 0.3 % samples above 1 ppm, 0.04 to 2.78 ppm for T-2 toxin with 25 % positive samples and 2.8 % samples above 1 ppm, and 0.003 to 0.06 for DAS with 21.6 % positive samples. In silages, no T-2 toxin was detectable. Isolated Fusarium strains were checked tor the ability to produce the mycotoxins zearalenone, T-2 toxin and DAS in culture. Most of the strains were positive for at least one of the toxins. Address: University of Bonn, Department of Agricultural and Food Microbiology, Meckenheimer Allee 168, D-53115 Bonn, Germany.

Natural Occurrence of Ochratoxin A and Citrinin in Food Stuffs in Egypt El-Sayed AM Abd Alla Mycotoxin Research 1996, Vol. 12 (1), 41-44

Natural occurrence of ochratoxin A (OA) and citrinin in cereals (274 samples) and animal tissues (250 samples) have been investigated during a period of more than 2 years. OA was found in cereals and animal tissues while citrinin was found in cereals only. The highest level of OA (up to 80.0 μg/kg) was found in yellow corn, 52.8 % of contaminated samples while respectively 55.9 % and 39.4 % of barley and rice samples were contaminated with citrinin, with the highest level up to 100.0 and 27.92 μg/kg for barley and rice respectively. The frequent contamination of animal kidney with OA (28 %

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positive out of 150 tested) average concentration 12.33 μg/kg. 2 % of liver and 4 % of muscles tissue were observed. Address: Mycotoxins Lab - National Research Centre Dokki – Cairo, Egypt.

Distribution of Trichothecene Mycotoxins in Maize Ears Infected with F. graminearum and F. crookwellense J Grabarkiewicz-Szczesna, E Foremska, and P Golinski Mycotoxin Research 1996, Vol. 12 (1), 45-50

Maize ears naturally infected with F. graminearum (6 samples) and F. crookwellense (6 samples) were collected in 1990 and 1991. All samples exhibited symptoms of severe ear rot with high percentage of Fusarium damaged kernels. Thousand kernels weight ranged 108.5–235.0 g and was significantly lower than in control, not infested kernels (330 g). All ears were contaminated with Fusarium toxins but DON and NIV were the most frequently analyzed metabolites. Distribution of trichothecene mycotoxins in different parts of ear as well as toxicity of extracts to Artemia salina is reported in this paper. Address: Department of Chemistry, Agricultural University of Pozna, ul Wojska Polskiego 75, 60-625 Pozna, Poland.

Secondary Metabolites Produced by Alternaria infectoria and Their Use as Chemotaxonomic Markers Birgitte Andersen and Ulf Thrane Mycotoxin Research 1996, Vol. 12 (2), 54-60

Twenty-one isolates of Alternaria infectoria have been screened for their production of secondary metabolites for chemotaxonomic characterization. For comparison isolates of A. tenuissima and Stemphylium sarciniforme were also screened. A. infectoria and A. tenuissima had two unknown metabolites and none known metabolites in common. A. infectoria had two other unknown metabolites in common with S. sarciniforme. The positions of six unknown metabolites in the metabolite profile of A. infectoria have been determined. UV-spectra and retention time indices of these six metabolites are given. Address: Department of Biotechnology, Building 221, Technical University of Denmark, DK-2800 Lyngby, Denmark.

Cyclopiazonic Acid and Aflatoxins Production by Aspergillus flavus Isolated from Argentinian Corn 1,2 3,4 2,5 6 1 6 SL Resnik , HHL González , AM Pacin , M Viora , GM Caballero , and EG Gros

Mycotoxin Research 1996, Vol. 12 (2), 61-66

Thirty-four isolates of Aspergillus flavus obtained from the main Argentinian corn production area were tested for their ability to produce both cyclopiazonic acid (CPA) on corn and on liquid media and aflatoxins on corn. Aflatoxins and CPA were quantified by comparison with standards. The last one was confirmed by mass spectrometry. All but one of the isolates produced CPA on liquid medium in a range between 3120 to 62500 μg/kg, 27/34 isolates produced CPA on corn at levels ranging from 833 to 10000 μg/kg and 5/34 isolates produced aflatoxin B1 in a range between 29 to 115 μg/kg. According to these findings, the percentage of Aspergillus flavus isolates with CPA production ability and their levels of CPA production were higher than the observed elsewhere. It was observed significant differences (p<0.01) between CPA production on corn (median: 1761 μg/Kg) and in liquid medium 241

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(median: 27950 μg/Kg). These data represent the first report of the co-production of CPA and aflatoxin B1 by isolates of Aspergilus flavus obtained from corn in Argentina. Addresses: 1, Facultad de Ciencias Exactas y Naturales, UBA, Buenos Aires, Argentina; 2, Comisión de Investigaciones Científicas de la Provincia de Buenos Aires, Buenos Aires, Argentina ; 3, Facultad de Ingeniería, UBA, Buenos Aires, Argentina; 4, Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina; 5, Universidad Nacional de Luján-Centro de Investigaciones en Micotoxinas, Argentina; 6, Facultad de Veterinaria, UBA, Buenos Aires, Argentina.

Toxicity of Extracts of Barley Kernels Inoculated with Fusarium spp. to Artemia salina 1 2 1 1 E. Foremska , I. Kiecana , J. Perkowski , and P. Golinski

Mycotoxin Research 1996, Vol. 12 (2), 67-72

Toxicity to A. salina of the Fusarium metabolites: deoxynivalenol (DON), its acetylated derivatives (3- and 15-AcDON), zearalenone (ZON), neosolaniol (NEO), nivalenol (NIV), T-2, HT-2 toxins, has been examined and compared with toxicity of extracts of barley kernels (8 cultivars and 4 lines) inoculated with Fusarium culmorum, F. graminearum and F. sporotrichioides respectively. Estimated LC50 values were expressed as relative toxicity (RT) in mg DON/kg for samples inoculated with F. culmorum, F. graminearum or in mg T-2/kg for F. sporotrichioides inoculations. Toxicity of extracts of the same genotype/line kernels was compared among different pathogens used for inoculation and differences in Fusarium head blight susceptibility of different genotypes/lines inoculated with the same Fusarium strain were found. Significant correlation between toxicity of extracts (LC50, RT) and toxic metabolites concentration was found (r = 0.82; P = 0.01). Bioassays with A. salina offer a fast, easy and inexpensive method to examine cereal genotypes susceptibility to Fusarium head blight and mycotoxins accumulation in kernels. Addresses: 1, Department of Chemistry Agricultural University of Pozna, ul. Wojska Polskiego 75, 60-625 Pozna, Poland; 2, Department of Phytopathology, Agricultural University of Lublin, Poland.

Destruction of Aflatoxins B1 and G1 in Bread Making HA Amra, SAZ Mahmoud, AH Taha, and MA El-Azab Mycotoxin Research 1996, Vol. 12 (2), 73-78

The effect of processing steps as well preservatives used in French bread making namely propionic acid and/or potassium sorbate (0.2 %) on the destruction of aflatoxins B1 and G1 was studied. Mixing and baking processes showed marked destruction of aflatoxins B1 and G1; being 71.2 % and 52.5 % for aflatoxin B1 after mixing and baking steps, while reaching 73.9 % and 54.5 % for aflatoxin G1. Fermentation step caused additional 15.3 % and 15.0 % destruction of aflatoxin B1 and G1. On the other hand, aflatoxin B1 destruction was 79.2 % and 50.7 % when propionic acid was used and 75.3 and 56.7% in the presence of potassium sorbate and after mixing and baking steps respectively. Concerning aflatoxins G1 it was found that mixing and baking steps showed destruction of 81.9 % and 53.4 % in the presence of propionic acid and 75.1 and 49.4 % in the presence of potassium sorbate in this respective order. Generally, it can be concluded that using propionic acid as preservative appeared to be more effective on the destruction of aflatoxins B1 and G1 than potassium sorbate in French bread making. Address: Dairy and Food Technology Dep., National Research Centre, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.

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Purification and Quantification of Nivalenol R Hedman and H Pettersson Mycotoxin Research 1996, Vol. 12 (2), 79-90

A mean of 2.79 g fusarenon-X (FUS-X)/L was produced in cultures of Fusarium sp Fn-2B on SSA, a synthetic medium containing sucrose and asparagine. When culturing Fn-2B on several other media, or using other Fusarium strains, much lower concentrations of FUS-X or nivalenol (NIV) were obtained. The SSA-incubations with Fn-2B in an optimal volume of 100 mL were stopped just before reaching maximal FUS-X concentration and the start of conversion to NIV. FUS-X was extracted from the 22 days old cultures and partially purified on a silica gel column. It was then hydrolysed to NIV, which was rechromatographed on silica and crystallized. The purity and the chemical and physical properties of the NIV produced were investigated. Potential sources of error when quantifying NIV were also studied. Address: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, 750 07 Uppsala, Sweden.

The Respiration-Impairing Effect of Rubroskyrin, a Toxic Metabolite of Penicillium islandicum Sopp, on Isolated Mitochondria Satoru Mori1, Yukiko Sugihara2, Akira Kitagawa2, Kiyoshi Kawai2, Yoshinori Nozawa1, and Yukio Ogihara3 Mycotoxin Research 1996, Vol. 12 (2), 91-98

The toxic effect of rubroskyrin, a modified bis-anthraquinone pigment from Penicillium islandicum Sopp, on mitochondrial respiration has been studied by using isolated rat liver mitochondria, comparing with those of luteoskyrin and rugulosin which are well known islandicum toxins. lt was found that rubroskyrin exerted an uncoupling effect on mitochondrial respiration, abating the respiratory control ratio (RCR) by a dose dependent manner (UD50: 10 μM) and markedly depressed state 3 respiration at high concentrations, whereas such respiration-impairing effects of luteoskyrin and rugulosin were not detected at the concentrations tested (max. 35 μM). The involvement of redox-reaction was not detected in the uncoupling effect of the quinone pigment, rubroskyrin. Spectroscopic study at various pHs revealed that rubroskyrin possessed a pK value within the range of physiological pH, suggesting that rubroskyrin is a weak acid-type uncoupler capable of conducting protons across mitochondrial inner membrane. Addresses: 1, Department of Biochemistry, Gifu University School of Medicine, Gifu 500, Japan; 2, Department of Nutrition, Faculty of Wellness, Chukyo Women‘s University, Ohbu 474, Aichi, Japan; 3, Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya 467, Japan.

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Fate of Aflatoxins B1 and G1 Residues During Biscuit Processing With or Without Leavening Agents HA Amra, SAZ Mahmoud, AH Taha, and MA El-Azab Mycotoxin Research 1996, Vol. 12 (2), 99-104

Effect of biscuit processing on the destruction of aflatoxins B1 and G1 with and/or without some commonly leavening agents used namely sodium bicarbonate, ammonium bicarbonate and sodium bisulfite and sodium chloride. lt was found that mixing step reduced the concentration of aflatoxins B1 and G1 by 80.7 % and 82.7 %, while the effect of baking step being 28.9 % and 21.5 %. The effect of mixing was found to be more pronounced than that baking step. The highest destruction effect on aflatoxin B1 was observed by adding a mixture composed of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium chloride, sodium bisulfite, ammonium bicarbonate and/or sodium bicarbonate alone, where the reduction values of toxin after mixing were 93.4, 91.9, 91.7, 88.8 and 86.6 % respectively, while the baking effect ranged 17.2 to 34.5 % in the presence of different leaving agents added. Concerning aflatoxin G1, the highest destructive effect of toxin was adsorbed by adding a mixture of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium bisulfite, sodium chloride, ammonium bicarbonate and/or sodium bicarbonate alone since the destruction values of such toxin after mixing were 96.2 %, 92.8 %, 92.6 %, 89.0 % and 87.7 % respectively, while the baking effect ranged 20.9 to 34.5 % in all leavening agents added. Address: Dairy and Food Technology Dep., National Research Centre, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.

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