Brine Shrimp ( Artemia salina L.) Larvae as a Screening System for Fungal Toxins

Abstract

Concentrations resulting in 50% mortality, determined with brine shrimp ( Artemia salina L.) larvae exposed to known mycotoxins for 16 hr, were (μg/ml): aflatoxin G 1 , 1.3; diacetoxyscirpenol, 0.47; gliotoxin, 3.5; ochratoxin A, 10.1; and sterigmatocystin, 0.54. 4-Acetamido-4-hydroxy-2-butenoic acid γ-lactone gave no mortality at 10 μg/ml. Used as a screening system involving discs saturated with solutions of known mycotoxins, the larvae were relatively sensitive to aflatoxin B 1 , diacetoxyscirpenol, gliotoxin, kojic acid, ochratoxin A, rubratoxin B, sterigmatocystin, stemphone, and T-2 toxin. Quantities of 0.2 to 2 μg/disc caused detectable mortality. The larvae were only moderately sensitive to citrinin, patulin, penicillic acid, and zearalenone which were detectable at 10 to 20 μg/disc. They were relatively insensitive to griseofulvin, luteoskyrin, oxalic acid, and β-nitropropionic acid. The disc screening method indicated that 27 out of 70 fungal isolates from foods and feeds grown in liquid or solid media produced chloroform-extractable toxic material. Examination of toxic extracts by thin-layer chromatography for 17 known mycotoxins showed that the toxicity of eight isolates could be attributed to aflatoxin B 1 and B 2 , kojic acid, zearalenone, T-2 toxin, or ochratoxin A. Nine out of 32 of these fungal isolates grown in four liquid media yielded toxic culture filtrates from at least one medium. Chemical tests for kojic, oxalic, and β-nitropropionic acids showed the presence of one or two of these compounds in filtrates of seven of these nine isolates.