Abstract. The lighting regime significantly impacts poultry production, reproductive performance, health and
welfare. This study aimed to test the effect of the intermittent light (IL) regime on reproductive organs and
hormones, semen quality, and behavioral performance. Thus, 270 Rhode Island Red hens aged 20 weeks were
distributed among three groups of six replicates and 15 birds each, housed in floor pens. The first group was used
as a control (C) and was exposed to constant light for 16 h d?1, while birds in other groups were exposed to IL
for 20 min h?1 (T20) and 40 min h?1 (T40) during the 16 h light period. The outcomes were that follicle number
was higher for T20 compared to T40 but not the controls, while T40 is different from T20 but not the controls.
The same is true for testosterone. The sperm concentration is lower for T40 compared to the controls, with no
difference between T20 and the controls. Body temperature was not different among groups. Conversely, differences
were not noticed for leg alterations; plumage conditions; and relative ovary, oviduct, and/or testes weight
and hatchability. Thus, the IL T20 program should be further investigated as a lighting regimen for managing
Rhode Island Red laying hens for stimulating follicle number and testosterone without negatively impacting the
physiological response and health traits. From a practical point of view, the IL schedule of 20 min h?1 during
20–36 weeks of age can be economically viable due to saving 66% of the light cost.