Experiments were conducted to determine if a nocturnal rise in serum melatonin occurs in prepubertal gilts and whether acute exposure of gilts to light during the dark period abruptly reduces serum concentrations of melatonin. In experiment 1, 12 prepubertal crossbred gilts (Duroc x Hampshire x Chester White x Yorkshire) weighing 96.4 + 1.3 kg at 5.1 + .1 mo of age were housed in an LD cycle of 10:14. Following a 3-wk acclimation period, blood samples were drawn at 1-hr intervals from indwelling jugular catheters. Serum concentrations of melatonin were similar (P greater than .05) among blood samples collected during light and dark periods. In experiment 2, serum concentrations of melatonin did not change (P greater than .05) when gilts were abruptly exposed to light during the normal dark period. In experiment 3, serum concentrations of melatonin were similar (P greater than .05) in blood samples collected at 2-hr intervals under 700 lux of light or in total darkness from gilts maintained in either LD 9:15 or LD 24:0. Data from experiment 4 demonstrated that serum melatonin could be detected in nighttime samples if exogenous melatonin was ingested by gilts at night. Together, these experiments clearly indicate that prepubertal gilts do not exhibit a nocturnal rise in serum melatonin when maintained under short daylengths (10L:14D or 9L:15D), and serum melatonin concentrations are unaffected by abrupt changes in light/dark conditions.