Sterigmatocystin and Other Mycotoxins Produced by <I

711

Sterigmatocystin and other Mycotoxins Produced by Aspergillus Species NORMAN D. DAVIS Department ofBotany, Plant Pathology and Microbiology, Agricultural Experiment Station, Auburn University, Auburn, Alabama 36849

(Received for publication October 24, 1980) Downloaded from http://meridian.allenpress.com/jfp/article-pdf/44/9/711/1654452/0362-028x-44_9_711.pdf by guest on 30 September 2021

ABSTRACT The aflatoxins, ochratoxins and sterigmatocystin are the principal mycotoxins of Aspergillus spp. that are known to occur naturally in levels sufficient to be regarded as significant hazards to animal and human health. Adequate research has OH not been done to allow generalizations concerning the other 27 or more mycotoxins of Aspergillus spp. However, the widespread and frequent occurrence of toxigenic isolates of these fungi in feed and food materials indicates that concern is justified, and these fungi and their mycotoxins must be considered hazards until proven otherwise.

0 0

A recent text on food and beverage mycology (J) lists more than 40 Aspergillus spp. that are important to the food and beverage industry. At least 10 of these can Sterigmatocystin produce one or more of 30 mycotoxins. Except for Figure 1. The structure of sterigmatocystin. aflatoxin, ochratoxin, and perhaps sterigmatocystin, insufficient research has been done to determine whether xanthone and the pigments an anthraquinone ring fused these Aspergillus spp. and their mycotoxins constitute a to the furan system. significant health hazard to animals and man. Cole (,3), Biological effects ofsterigmatocystin on reviewing the chemistry of the Aspergillus toxins, The toxic effects of sterigmatocystin are much the stated that all of them should be considered hazards to same as those of aflatoxin B1 (25). Thus the compound is human and animal health until proven otherwise. This is viewed as a potent toxin, carcinogen, mutagen and certainly true of sterigmatocystin, which is a carcinogen, teratogen. Sterigmatocystin is generally less acutely toxic and of ochratoxin A, which is a naturally occurring than aflatoxin B1 (Table 1). The acute toxicity varies with nephrotoxin (2). Whether other Aspergillus toxins are the mode of application, the animal species and other important environmental food toxicants cannot be stated factors. TheLD 50 varies from 32 rug/kg of body weight in unequivocally. Sufficient research has not been directed the monkey to 800 mg/kg in mice. It is generally more to these toxins to determine their frequency and level of potent when injected than when administered per os, natural occurrence. Thus this report will deal largely suggesting that it is absorbed with difficulty (25). with sterigmatocystin and ochratoxin A. Chronic symptoms of sterigmatocystin poisoning have been summarized (6.25), and include induction of STERIGMATOCYSTIN hepatomas in rats, pulmonary tumors in mice, renal Sterigmatocystin (Fig. 1) belongs to the furofuran lesions and prominent alterations in the liver and kidney group of mold metabolites that includes the aflatoxins, of African Green monkeys. Other effects noted include sterigmatocystins and some closely related pigments (26). myocardial necroses in the heart, cellular alterations These metabolites contain either an unsaturated such as loss of chromatin from nuclei and progressive 7,8-dihydrofurano (2,3~b) furan or the more fully reduced cellular degeneration accompanied by nucleolar frag 2,3,7,8-tetrahydrofuro(2,3)furan system (26). The afla mentation. Rats fed 5-10 ppm of sterigmatocystin for toxins contain a coumarin, the sterigmatocystins a 2 years showed a 90o/oincidence of liver tumors (16).

JOURNALOFFOODPROTECTION. VOL.44,SEPTEMBER 1981 712 DAVIS

TABLE 1. Acute toxicity ofaflatoxin and sterigmatocystin. TABLE 3. Natural occurrence ofsterigmatocystin (STG) and sterigmatocystin ·producing fungi. LD50 (mg/kg)

Species and routea Aflatoxin B1 Sterigmatocystin Substrate Contaminant Monkey, p.o. 2.2-7.8 32 Wheat STG Rat, p.o. 7 .2(M)-16(F) 166(M)-120(F) Barley Rat, i.p. 7 .2(M)-17 .9(F) 60-7S(M) Rice Chicken embryo 0.025 1-1g/embryo 14.9 fAg/embryo Green coffee beans Mouse, i.p. 9.0 800 Pecans Zebra fish (LC 5o) 0.58 f.iglml 0.24 f.ig/ml Various grains STG-producing fungi intraperitoneal Bread " Wheat flour Meat Fungi that produce sterigmatocystin Cheese

Fifi:een species of fungi have been reported to produce Grape juice Downloaded from http://meridian.allenpress.com/jfp/article-pdf/44/9/711/1654452/0362-028x-44_9_711.pdf by guest on 30 September 2021 sterigmatocystin or sterigmatocystin-like compounds (fable 2). The principal producer is Aspergillus versi on the direct observation of the brick red fluorescence of color, which also produces 5-methoxy-sterigmatocystin, sterigmatocystin, per se, these methods are relatively 6-demethylsterigmatocystin, dihydrosterigmatocystin, insensitive, and about 500 ng is the smallest amount and dihydrodemethylsterigmatocystin (3.4). A. versicolor detectable. TLC methods are made more sensitive by has been used to produce up to 8 g of sterigmatocystin converting the sterigmatocystin into more intensely per kg of maize meal + soytone, whereas in other fluorescent derivatives. Stack and Rodricks (19) reported investigations A. versicolor produced up to 1.3 mg of a method for analysis of grains in which the toxin is sterigmatocystin per gram of mycelium (4). extracted with acetonitrile-water, defatted with hexane and partitioned into chloroform. The extract is then TABLE 2. Fungi that produce sterigmatocystin(s). partitioned on a silica column and sterigmatocystin is Aspergillus versicolor A. parasiticus eluted from the column and quantitated on a TLC plate. A. amstelodami A. quadrilineatus Quantitation is made more sensitive by spraying the A. aurantio·brunneus A. regulosus plate with A1Cl 3 and heating to convert sterigmatocystin A. chevalieri A. ruber into an intensely-fluorescing, yellow-green derivative. A.flavus A. sydowi Other methods for analysis of sterigmatocystin have A. nidulans A. ustus been developed. One method, that uses gel permeation Penicillium luteum Drechslera species followed by high pressure liquid chromatography, has Bipolaris sorokiniana been developed for analysis of corn and oats for sterigmatocystin (20). Several multiple mycotoxin analy Natural occurrence of sterigmatocystin and sterigmato tical methods include analysis for sterigmatocystin cystin-producingjimgi (6.10,17,23,27}. A method by Whidden et al. (27} is Toxigenic sterigmatocystin-producing fungi have been discussed under analytical methods for ochratoxin. found in a wide variety of foods and food materials OCHRATOXIN A (fable 3), e.g. A. versicolor has been isolated from most The ochratoxins are a group of mycotoxins that of the common grains, flour, bread, grape juice, meat consist of an isocoumarin moiety attached to a and cheese. Sterigmatocystin, per se, has been found as a phenylalanine moiety. Of nine or more ochratoxins that natural contaminant in relatively few instances (Table 3). are known to exist, only ochratoxin A (Fig. 2) is thought It has been found in severely molded wheat, barley, rice, to be of significance as a natural contaminant of food pecans and green coffee beans (24,26). Recently it has and feed (10). been reported as a contaminant of marihuana (12) and Biological ejfects o_f'ochratoxin A in-shell pecans (18). The ochratoxins cause damage in several species of Analytical methods for sterigmatocystin animals, including mice, rats. dogs, pigs, ducklings, The paucity of information on the natural occurrence hens, rainbow trouts and monkeys (10). Acute toxicity is of sterigmatocystin in foods and feeds is probably in about 0.2-0.34 mg/kg of body weight for chick embryos, large part due to the lack of sensitive methods suitable 2.1 mg/kg for day-old-chickens and approximately 1.88 for general screening for sterigmatocystin. Present mg/kg for day-old-ducklings (Table 4) (8). This is less methods of analysis are sensitive to 10-50 ppb, but are toxic generally than aflatoxin, but more toxic than

regarded as unsuitable since screening should be done at sterigmatocystin. Generally, LD 50 values range from 0.2 5 ppb or less (2). Most methods developed for analysis of to 54 mg/kg. sterigmatocystin rely on thin layer chromatography Chronically. ochratoxin A is primarily a nephrotoxin (fLC) techniques for quantitation by visual comparison (11). Because of the striking similarities between Balkan of the intensity of fluorescence of authentic standards of endemic nephropathy and ochratoxin A-induced porcine sterigmatocystin and sample extracts (26). When based nephropathy, ochratoxin A has been suggested as a

JOURNAL OF FOOD PROTECTION. VOL. 44. SEPTEMBER 1981 ASPERGILLUS-PRODUCED MYCOTOXINS 713

COOH 0 OH 0 I II documented for a number of agricultural commodities CH-NH-C (fable 6). Natural contamination has been observed in Bulgaria, Rumania, Yugoslavia, Sweden, Denmark and North America (3,10), and ochratoxin has been found in animal carcasses intended for distribution for human Cl consumption (22). Swine and poultry appear to be particularly susceptible to ochratoxicosis (7,13,22). Ochratoxin A TABLE 6. Natural occurrence ofochratoxin A a. Figure 2. The structure ofochratoxin A.

TABLE 4. Acute toxicity ofochratoxin A. Sample Level, ppb Corn 166 Organism Barley 127 Wheat 100 Duckling 1.88mg/kg Downloaded from http://meridian.allenpress.com/jfp/article-pdf/44/9/711/1654452/0362-028x-44_9_711.pdf by guest on 30 September 2021 White beans 2,100 Rat 20-22mg/kg Peanuts 4,900 Trout 4.67 mg/kg Barley/oats 27,500 Day-old-chicks 2.1 mg/kg "'"'uc:v~' 67,000 Chicken embryo 0.2-0.34 mg/kg Swine Zebra fish larvae 1. 7 11g/ml (LCso) aHarwig, 1974 (8).

Brine shrimp larvae 10 11g/ml (LC 5o) Ochratoxin-producing fungi have been found in corn, TABLE 5. Fungi that produce ochratoxinsa. rice, barley, oats, sorghum and wheat (Table 3) A. ochraceus P. viridicatum (3,8,10,21). They have also been isolated from peanuts, A. alliaceus P. commune pecans, Brazil nuts and cottonseed, and thus appear to A. melleus P. cyclopium be a general contaminant of oilseeds as well as grains. A. ostianus P. palitans Coffee beans, white beans, hops, ground pepper, adjuki A. petrakii P. purpurescens beans, dried fish, salted fish and citrus are among the A. sclerotiorum P. variabile miscellaneous products from which ochratoxin-produc A. sulphureus ing fungi have been isolated (3,8). aHarwig, 1974 (8). Levels of ochratoxin in natural products range from 100 ppb in wheat to 27,500 ppb in a severely molded possible cause of that disease (11). Other symptoms of mixture of barley and oats. Diseased kidneys were found ochratoxicosis include general impairment of the to contain 67,000 ppb ochratoxin A in one instance (8). immune system, hepatic lesions, renal lesions, renal In the laboratory, ochratoxin A has been produced by A. failure, impairment and necrosis, and in some species, ochraceus growing in laboratory media, on corn, fatty infiltration of the liver, or enteritis and visceral gout shredded wheat, soybeans and rice (21). Levels produced (7,8,13,29). Kidney impairment is the most general result ranged up to 239 mg/100 g of substrate (1). of ochratoxicosis, reported for most species tested; pale, swollen kidneys is the gross pathological symptom noted Analytical methods for ochratoxin A in most field outbreaks (2). Abortion in cows and The chemistry of the ochratoxins has been reviewed teratogenic effects in mice. rats and hamsters have also recentlv by Nesheim (14). He lists 152 references, mostly been reported (8,21,25). dealing with the qualitative and quantitative analysis of Fungi that produce ochratoxin A ochratoxins in natural products. He concludes that the multiplicity of methods described attests to the fact that Seven Aspergillus spp. and six Penicillium spp. have there is no single method adequate for general purposes. been identified as ochratoxin A producers (Table 5). The Association of Official Analytical Chemists (AOAC) Krogh (10) considers Penicillium viridicatum, Peni· Official Methods ofAnalysis (15) gives procedures for cillium palitans and Penicillium eye/opium to be the quantitative analysis of barley and green coffee ?eans for most important fungi involved in natural outbreaks of ochratoxin. Confirmation tests are also given for ochratoxicosis. However, toxigenic strains of Aspergillus ochratoxin A. In the official method for ochratoxin, the ochraceus are also frequently encountered in natural mycotoxin is extracted with aqueous phosphoric acid and products (21). P. viridicatum produces citrinin as well as chloroform, entrapped on a sodium bicarbonate ochratoxin A, and A. ochraceus produces penicillic acid diatomaceous earth column and, following cleanup of or mellein or hydroxymellein as well as ochratoxin A. the column with hexane and chloroform, the ochratoxins Thus synergism among those mycotoxins may exist. This are eluted with formic acid in chloroform. Ochratoxin A possibility has been little investigated and its importance is then quantitated on TLC plates by comparison of the in field outbreaks is not well understood. fluorescent intensitv of standards and extracts of Natural occurrence of ochratoxin A and ochratoxin· samples. Confirmation is via formation of ochratoxin producing fungi ethyl esters using BF 3 and cochromatography with The natural occurrence of ochratoxin A has been standards on TLC plates. Another useful technique is to

JOURNAL OFFOOJJ PROTECTION. VOL 44. SEPTEMBER 1981 714 DAVIS examine the TLC plates under long and short wave UV S. Durackova, Z., V. Betina, and P. Nemec. 1976. Systematic before and after exposure to fumes of ammonia. analysis of mycotoxins by thin-layer chromatography. J. Chrom Ochratoxin A fluoresces much darker blue following atog.ll6:141-154. 6. Hamasaki, T., andY. Hatsuda.l977. Sterigmatocystin and related exposure to ammonia. Unlike many other compounds, it compounds. pp. S97-607.ln J. V. Rodricks, C. W. Hesseltine, and can be observed under both long and short wave UV M. A. Mehlman (ed.) Mycotoxins in human and animal health. radiation. Sensitivities are as low as 2 ppb. Several Pathotox PubI. Park Forest South, Illinois. multimycotoxin methods include analysis for ochratoxin 7. Hamilton, P. B., W. E. Huff, J. R. Harris, and R. D. Wyatt. 1977. A (5,17,23,27). A scheme by Whidden et al. (27) involves Outbreaks of ochratoxicosis in poultry. Am. Soc. Microbial. Ann. Mtg.0-21. extraction of acidified corn with acetonitrile, defatting 8. Harwig, J. I974. Ochratoxin A and related metabolites. pp. with isooctane, partition on and subsequent elution from 345-367. In I.F.H. Purchase (ed.) Mycotoxins. Elsevier Science a silica column using chloroform-methanol (95:5). Publ. Co., New York. Ochratoxin A is quantitated by TLC methods as 9. Josefsson. B. G. E., and T. E. Moller. 1977. Screening method for previously described. This method includes analysis for the detection of atlatoxins, ochratoxins, patulin, sterigmatocystin,

and zearalenone in cereals. J. Assoc. Off. Anal. Chern. Downloaded from http://meridian.allenpress.com/jfp/article-pdf/44/9/711/1654452/0362-028x-44_9_711.pdf by guest on 30 September 2021 sterigmatocystin, zearalenone, patulin, penicillic acid, 60:1369-1371. oxchratoxin A, aflatoxin B1, diactoxyscirpenol and 10. Krogh, P. 1977. Ochratoxins pp. 489-498. In J. V. Rodnicks, C. W. rubratoxin B. Hesseltine and M. A. Mehlman (ed.) Mycotoxins in human and animal health. Pathotox Pub]. Park Forest South, Illinois. 11. Krogh, P., N.H. Axelsen, F. Elling, N. Gyrd-Hansen, B. Hald, J. OTHER MYCOTOXINS OF ASPERGILLUS SPECIES Hyldgaard-Jensen. A. E. Larsen, A. Madsen, H. P. Mortensen, More than 30 additional mycotoxins. produced by 10 T. Moller, 0. K. Petersen, U. Ravnskov, M. Rostgaard, and 0. or more Aspergillus spp. have been described (fable 7) Aalund. 1974. Experimental porcine nephropathy. Changes of (3,28). Many of these are potent mycotoxins; however, it renal function and structure induced by ochratoxin A-contam inated reed. Acta Pathol. Microbiol. Scand. Sect. A., Suppl. No. is not known if they are produced naturally in foods and 246:1-21. feeds by Aspergillus spp., or in amounts that might make 12. Llewellyn, G. C.. and C. E. O'Rear. 1978. A preliminary evalua them significant health hazards. tion of illicit marihuana (Cannabis sp.) for mycotoxins. Dev. Ind. Microbial. 19:319-323. TABLE 7. Miscellaneous Aspergillus toxinsa. 13. Nelson. G. H. 1979. Biological effects of mycotoxins (other than allatoxin) on swine. pp. 77-86. In Interactions of mycotoxins in Species Mycotoxin(s) animal production. National Academy of Science. 2101 Constitu A. candidus Citrinin, candidulin, kojic acid tion Avenue N.W .. Washington, D.C. 20418. 14. Nesheim, S. 1976. The ochratoxins and other related compounds. (3- nitropropionic acid pp. 276-295. In Mycotoxins and other fungal related food problems. A. clavatus Patulin, ascladiol, cytochalasin Ad. Chern. Ser. 149, Am. Chern. Soc., Washington, D.C. E 15. Official Methods of Analysis, 12 ed. 1975. AOAC, Arlington, VA. A.flavus Kojic acid, (3-nitropropionic Chapter 26. acid, aspergillic acid. aspertoxit;I, 16. Ohtsubo, K., M. Saito, H. Kimura, and 0. Tsuruta. 1978. High flavutoxin, oxalic acid incidence of hepatic tumours in rats fed moldy rice contaminated A.jumigatus Viriditoxin, gliotoxin, helvolic with Aspergillus versicolor containing sterigmatocystin. Food acid, fumagillin, terrein, Cosmet. Toxicol. 16:143-150. spinulosin 17. Roberts. B. A .. and D. S. P. Patterson. 1975. Detection of twelve A. chevalieri Gliotoxin, bovine toxin mycotoxins in mixed animal feedstuffs using a novel membrane cleanup procedure. I. Assoc. Off. Anal. Chern. 58:1178-1181. A. oryzae var. microsporis Maltoryzine 18. Schroeder, H. W., and H. Hein, Jr. 1977. Natural occurrence of A. niger Malformin, oxalic acid, kojic sterigmatocystin in in-shell pecans. Can. J. Microbial. 23:639-641. acid 19. Stack, M., and J. V. Rodricks. 1971. Methods for analysis and A. ochraceus Penicillic acid, mellein, chemical confirmation of sterigmatocystin. J. Assoc. Off. Anal. hydroxymellein Chern. 54:86-90. A. terreus Patulin, citrinin, gliotoxin, 20. Stack, N. E., S. Nesheim, N. L. Brown, and A. E. Pohland. 1976. terretoxin Determination of sterigmatocystin in corn and oats by gel permeation and high-pressure liquid chromatography. J. Assoc. a Tuite, 1977 (24) and Cole, 1976 (3). Off. Anal. Chern. 59:%6-970. 21. Steyn, P. S. 1971. Ochratoxin and other dihydroisocoumarins. pp. REFERENCES 179-205. In A. Ciegler, S. Kadis. and J. Ajl (ed.) Microbial toxins, Vol. VI. Fungal toxins. Academic Press, N.Y. I. Beuchat. L. R. (ed.). 1978. Food and beverage mycology. AVI 22. Stoloff. L. 1979. Mycotoxin residues in edible animal tissues. Pub!. Co., Inc .. Westport. Conn. pp. 157-166. In Interactions of mycotoxins in animal production. 2. CAST. 1979. Aflatoxin and other mycotoxins: an agricultural National Academy of Science. 2101 Constitution Avenue. N.W .. perspective. Council for Agricultural Science and Technology. Washington, D.C. 20418. Report No. 80.250 Memorial Union, Ames. Iowa 50011. 23. Stoloff. L.. S. Nesheim, L. Lin, J. V. Rodricks, M. Stack, and 3. Cole, R. J. 1976. Aspergillus toxins other than aflatoxin. pp. 68-89. A. D. Campbell. 1971. A multimycotoxin detection method for In J. V. Rodricks (ed.) Mycotoxins and other fungal related food atlatoxins. ochratoxins. zearalenone. sterigmatocystin, and pat problems. Adv. Chern. Ser. 149. Am. Chern. Soc., Washington, ulin. J. Assoc. Off. Anal. Chern. 54:91-97. D.C. 24. Tuite, J. 1977. The genus Aspergillus. pp. 21-39. T. D. Wyllie and 4. Detroy. R. W., E. B. Lillehoj, and A. Ciegler. 1971. Aflatoxin and L. G. Morehouse (ed.) Mycotoxic fungi. mycotoxins and myco related compounds. pp. 3-178. In A. Ciegler, S. Kadis and S. J. Ajl toxicoses. Vol. l. Marcel Dekker, N.Y. (ed.) Microbial toxins, Vol. VI. Academic Press N.Y. 25. Ueno. Y .. and I. Ueno. 1978. Toxicology and biochemistry of con't. p. 722

JOURNAL OF FOOD PROTECTION. VOL. 44. SEPTEMBER 1981 722 COLE

5. Biichi, G., K. C. Luk, B. Kobbe, and J. M. Townsend. 1977. Four 22. Mantle, P. G .. P. H. Mortimer. and E. P. White. 1977. Mycotoxic new mycotoxins of Aspergillus clavatus related to tryptoquivaline. tremorgens of Claviceps paspali and Penicillium eye/opium: a J. Org. Chern. 42:244-246. comparative study of effects on sheep and cattle in relation to 6. Cardillo, R., C. Fuganti, G. Gatti, D. Chiringelli, and P. natural staggers syndromes. Res. Vet. Sci. 24:49-56. Grasselli. 1974. Molecular structure of cryptoechinulin A, a new 23. Martins, C. 1929. Etudes experimentales sur !'Aspergillus metabolite of Aspergillus amstelodami, isolated during investiga fumigatus. C. R. Soc. Bioi. (Paris) 100:525-526. tions on echinulin biosynthesis. Tet. Lett. 36:3163-3166. 24. Moreau, C.. and M. Moreau. 1960. Un danger pour le betail 7. Ciegler, A., R. F. Vesonder. and R. J. Cole. 1976. Tremorgenic nourri de plantules fourrageres cultivees engermoir; Ia polution mycotoxins. Adv. Chern. 149,163-177. d'une moisissure toxique, !'Aspergillus clavatus, cause des 8. Clardy, J., J. P. Springer, G. Biichi, K. Matsuo, and R. accidents mortels. C. R. Acad. Agric. Fr. 46:441-445. Wightman. 1975. Tryptoquivaline and tryptoquivalone, two 25. Raper, K. B .. and D. I. Fennell. 1965. The genus aspergillus. tremorgenic metabolites of Aspergillus clavatus. J. Am. Chern. Williams & Wilkins Co., Baltimore. Soc. 97:663-665. 26. Richard, J. L., and L. H. Arp. Natural occurrence of the 9. Cole, R. J. 1977. Tremorgenic mycotoxins. In J. V. Rodricks, C. mycotoxin penitrem A in moldy cream cheese. Mycopathologia W. Hesseltine, and M.A. Mehlman (eds.) Mycotoxins in human (In press.) and animal health. Pathotox Publishers, Inc., Park Forest South, 27. Rutquist, L. 1965. Studies on Aspergillus fumigatus; toxin Ill. production by different strains and serological comparison of the Downloaded from http://meridian.allenpress.com/jfp/article-pdf/44/9/711/1654452/0362-028x-44_9_711.pdf by guest on 30 September 2021 10. Cole, R. J., J. W. Kirksey, J. W. Dorner, D. M. Wilson, J. C. strains. Acta Vet. Scand. 6. 224-233. Johnson. Jr., D. Bedell, J. P. Springer, K. K. Chexal. J. Clardy, 28. Smith. D. F .. and G. P. Lynch. 1973. Aspergillus .fumigatus in and R. H. Cox. 1977. Mycotoxins produced by Aspergillus samples ofmo1dy silage. J. Dairy Sci. 56,828-829. fumigatus isolated from silage. Ann. Nutr. Alim. 31:685-691. 29. Springer, J.P. 1979. The absolute configuration of nortrytoquiva 11. Cole. R. J., J. W. Kirksey. J. W. Dorner, D. M. Wilson, J. C. line. Tet. Lett. 43:339-342. Johnson, Jr., A. N. Johnson, D. M. Bedell, J. P. Springer. K. K. 30. Steyn, P. S. 1971. Austamide, a new toxic metabolite from Chexal, J. C. Clardy, and R. H. Cox. 1977. Mycotoxins produced Aspergillus ustus. Tet. Lert. 36:3331-3334. by Aspergillus .fumigarus species isolated from molded silage. J. 31. Whitehair, C. K .. H. C. Young. Jr., M. E. Gibson, and G. E. Agric. Food Chern. 25:826-830. Short. 1951. A nervous disturbance in cattle caused by a toxic 12. Di Menna, M. E .. P. G. Mantle. and P. H. Mortimer. 1976. substance associated with mature Bermuda grass. Feeders' Day Experimental production of a staggers syndrome in ruminants by Reports pp. 57-59. a tremorgenic Penicillium from soil. N. Z. Vet. J. 24:45-46. 32. Wicklow, D .. and R. J. Cole. 1980. The distribution of aflatoxin. 13. Gallagher. R. T .. R. G. Keogh, G. C. M. Latch, and C. S. W. cyclopiazonic acid, aflatrem and dihydroxyatlavinine between Reid. 1977. The role offungal tremorgens in ryegrass staggers. N. sclerotia] and non-sclerotia! producing isolates of A. jlavus. Z. J. Agric. 20:431-440. Unpublished data. 14. Gallagher, R. T .. J. Clardy, and B. J. Wilson. 1980. Atlatrem. a 33. Wilson, B. J., and C. H. Wilson. 1964. Toxin from Aspergillus tremorgenic toxin fromAspergillusflavus. Tet. Lett. 21:239. .flavus: production on food materials of a substance causing 15. Gallagher, R. T., G. C. M. Latch, and R. G. Keogh. 1980. The tremors in mice. Science 144:177-178. janthitrems: Fluorescent tremorgenic toxins produced by 34. Wilson, B. J., C. H. Wilson, and A. W. Hayes. 1968. Tremorgenic Penicillium janthinellum isolates from ryegrass pastures. Appl. toxin from Penicillium cyclopium grown on food materials. Environ. Microbiol. 39:272-273. Nature 220:77-78. 16. Glinsukon, T .. S. S. Yuan, R. Wightman. V. Kitaura, G. Biichi. 35. Yamazaki, M .. H. Fujimoto, T. Akiyama, U. Sankawa, and Y. R. C. Shank, G. N. Wogan, and C. M. Christensen. 1974. litaka. 1975. Crystal structure and absolute configuration of Isolation and purification of cytochalasin E and two tremorgens fumitremorgin B, a tremorgenic toxin from Aspergillusfumigatus from Aspergillus clavatus. Plant Foods Man I :113-119. Fres. Tet. Lett. 14:27-28. 17. Hopkirk. C. S. M. 1936. Paspalum staggers. N. Z. J. Agric. 36. Yamazaki, M .. H. Fujimoto, and T. Kawasaki. 1975. The 105-!08. structure of a tremorgenic metabolite from Aspergillus fumigarus 18. Jacquet, J.. P. Boutibonnes. and J. P. Cicile. 1963. Observations Fres., fumitremorgin A. Tet. Lett. 14:1241-1244. sur Ia toxicite d'Aspergillus clavatus pour les animaux. Bull. 37. Yamazaki. M .. H. Fujimoto, and E. Okuyama. 1976. Structure Acad. Vet. Fr. 36:199-207. determination of six tryptoquivaline-related metabolites from 19. Kellerman, T.S .. J. G. Pienaar, G. C. A. Vander WestHuizen. L. Aspergillusfumigatus. Tet. Lett. 15:2861-2864. A. P. Anderson. and T. W. Naude. 1976. A highly fatal 38. Yamazaki. M., H. Fujimoto. and E. Okuyama. 1977. Structure of tremorgenic mycotoxicosis of cattle caused by Aspergillus tryptoquivaline C (FTC) and D (FTD), novel fungal metabolites clavatus. Onderstepoort.J. Vet. Res.43:147-154. from Aspergillus fumigatus. Chern. Pharmacol. BulL 25:2554- 20. Keogh. R. G. 1972. Induction and prevention ofryegrass staggers 2560. in grazing sheep. N. Z. J. Exp. Agric. I :55-57. 39. Yamazaki. M .. H. Fujimoto. and E. Okuyama. 1978. Structure 21. Ling, K. H .. C. K. Yang, and F. Peng. 1979. Territrems, determination of six fungal metabolites. tryptoquivaline E. F. G, tremorgenic mycotoxins of Aspergillus terreus. Appl. Environ. H. I. and J from Aspergillus fumigatus. Chern. Pharmacol. Bull. Microbiol. 37:355-357. 26:111-117.

Davis, con·t.fromp. 7!4

mycotoxins. pp. 107-155. InK. Uraguchi and M. Yamazaki (ed.) Toxicology, biochemistry and pathology of mycotoxins. John Wiley 28. Wilson, B. J. 1971. Miscellaneous Aspergillus toxins. pp. 207-295. & Sons, New York. In A. Ciegler. S. Kadis, and S. Ajl (ed.) Microbial Toxins. Vol. VI. 26. Vander Watt, J. J. 1974. Sterigmatocystin. pp. 370-382. In I.F.H. Academic Press. N.Y. Purchase (ed.) Mycotoxins. Elsevier Scientific Pub!. Co., New York. 29. Wyatt. R. D. 1979. Biological effects of mycotoxins (other than 27. Whidden, M.P .. N.D. Davis, and U. L. Diener. 1980. A method aflatoxin) on poultry. pp. 87-95. 1n Interaction~ of mycotoxins to detect rubratoxin B and seven other mycotoxins in corn. J. in animal production. National Academy of Science. 2101 Agric. Food Chem. 28. Constitution Avenue, N.W .. Washington, D.C. 20418.

JOURNAL OF FOOD PROTECTION. VOL. 44, SEPTEMBER 1981