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The estimation of melatonin in bovine plasma by radioimmunoassay is described and some aspects of specificity are discussed. Mean values for individual heifers ranged from 0·025 to 0·25 nmol/l during the day and were significantly (P<0·01) raised in each animal to mean values of 0·19 to 1·41 nmol/l at night. The hormonal changes after cloprostenol injections were not affected by the time of day at which the heifers were treated.