Rabbit Skin Test for Estimation of T-2 Toxin and Other Skin-Irritating Toxins in Contaminated Corn

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Abstract
A rabbit bioassay test has been developed which can be used for screening large numbers of field samples of corn for T-2 and other toxins. Two μl each of the control solutions, 0.005–0.12 μg standards, and about 12 samples in ethyl acetate were applied simultaneously to the closely clipped skin of a 2–3 kg young rabbit. The skin reactions were read at 24, 48, and 72 hr. The prominent features of the reactions were erythema, edema, and necrosis. The T-2 toxin equivalences of the samples were estimated by the degree of the skin reactions caused by the standards. The method is reliable to at least 0.01 μg/test or 50 ppb. Graded response to the toxin is dependent upon the use of sensitive animals and properly diluted samples and standards. Rabbits were more sensitive than weanling rats or young guinea pigs, and the method showed better reproducibility with rabbits. Citrinin and zearalenone (F-2) showed negative skin reactions when applied at a level of 20 μg/skin site. They also showed poor skin reactions when administered by intradermal injection into rabbit skin. However, all 3 mycotoxins showed distinct skin reactions in guinea pigs by the intradermal route. The test was as sensitive as the topical test using rabbits with T-2 toxin. The prominent feature of the skin reaction was induration.

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