Transiently evoked otoacoustic emissions (TEOAEs) have been widely used in universal newborn hearing screening programs. Although there is consensus with regard to the avoidance of early screening, especially during the first hours after birth, the optimum testing day is not yet unanimously accepted. The aim of the present study was to compare the ‘pass-refer’ results between 4 groups of newborns tested during the 4 postbirth days and determine the most appropriate day for assessing newborn hearing. Our results suggest that, although TEOAEs can be recorded in very high rates from the first 24 h of life, ‘refer’ scores are lower on the third and fourth days after birth. It may be thus concluded that the optimum time of assessing newborn hearing in universal hearing screening programs seems to be the third or fourth postbirth day, provided that other social or financial reasons do not suggest an earlier discharge from the hospital.