Diagnosis of Japanese Star Anise ( Illicium anisatum L.) Poisoning in a Holstein Heifer by Morphological Observation, PCR Method and LC/MS Analysis

Abstract

In September 2009, two of four Holstein heifers in a farm in Saitama Prefecture showed astasia and convulsions, and one died. Two days earlier, the owner of the farm had provided new bedding for the heifers by cutting branches from trees near the paddock. Partially digested leaves were found in the rumen of the dead heifer, and were identified as Japanese star anise (Illicium anisatum L.) by morphological observation, the PCR-RFLP method, and phylogenetic analysis. Anisatin, the toxic compound naturally occurring in Japanese star anise, was also' detected in the ruminal content and serum by liquid chromatography mass spectrometry (LC/MS). On the basis of these findings, the death of the heifer was determined to be a result of Japanese star anise poisoning. A combination of morphological, genetic, and chemical methods is available for diagnosis of plant poisoning.