Clarifying plasma melatonin profiles in domestic pigs: a critical and comparative evaluation of two radioimmunoassay systems

The results of a direct radioimmunoassay (RIA) for porcine plasma melatonin, suggesting a relationship among plasma melatonin, feed intake and photoperiod, were investigated by comparison with the results of an extracted RIA. These findings were further examined by analysis of a small number of samples using gas chromatography-mass spectrometry (GCMS). The results identified inadequacies in the specificity of the direct RIA and failed to provide evidence for an effect of feed intake on melatonin secretion. Instead, it appeared that feed restriction exacerbated nonspecificity in the direct RIA. The cause of nonspecificity, and its effect on analysis of daily plasma melatonin profiles, were examined. The plasma component(s) responsible for nonspecificity was (were) not identified. However, the results suggest that characteristics of the antiserum used in the direct RIA are involved in the mechanism by which nonspecificity is induced. Results obtained using the extracted assay revealed that plasma melatonin concentrations in prepubertal gilts and pregnant sows exhibit a diurnal pattern, in which concentrations are low during daylight and modestly increased during darkness. Using the direct RIA, the same profiles exhibited highly variable melatonin concentrations showing little association with the light-dark cycle. Thus, assay specificity was identified as a factor contributing to inconsistencies in the literature describing plasma melatonin in the domestic pig and the importance of rigorous validation of RIAs was demonstrated. Furthermore, the results indicate that plasma melatonin concentrations in domestic pigs, as in other mammalian species, are entrained by the light-dark cycle