One of the critical factors in assessing maize lines for resistance to Sesamia calamistis is the number of larvae/plant used for infestation. Artificial infestation of maize plants with two and five larvae/plant resulted in 29.1 and 39.5% stand loss (dead hearts and stalk breakage), while 10 and 15 larvae/plant resulted in 52 and 72.9% stand loss, respectively. For a satisfactory artificial infestation in maize screening trials against the maize stem borer in Nigeria, a population level of 10–15 larvae/plant is recommended.