LAUSR

&NA; We hypothesized that performance and bone mineralization of 2 broiler lines will benefit from increasing vitamin D (vitD) supplementation above current commercial levels and by partial substitution of D3 by 25‐OH‐D3. Male Ross 308 and 708 chicks (n = 576), were offered diets with low (LD; 1,000), medium (MD; 4,000) or high levels of D3 (HD; 7,000 IU/kg), and medium levels of vitD where the majority of D3 was substituted by 25‐OH‐D3 (25MD; 1,000 D3+3,000 25‐OH‐D3 IU/kg). Performance was measured at the end of starter (day 10), grower (day 24), and finisher periods (day 38). Three birds per pen were dissected at the end of each period to assess tibia and femur ash percentage (%), ash weight, bone breaking strength (BBS), and serum levels of 25‐OH‐D3. Remaining birds were gait scored (GS) at day 37 of age. Genotype and diet did not interact for any trait, whilst performance was not affected by diet. Ross 708 had lower body weight (P < 0.005), higher feed conversion ratio over the grower period (P < 0.05), similar levels of 25‐OH‐D3, but higher GS (P < 0.05) than Ross 308. Serum 25‐OH‐D3 levels were affected by diet at the end of the starter and grower periods (P < 0.05), being lowest for LD and highest for 25MD. Diet affected GS (P < 0.01), being higher in LD than 25MD. Femur ash % was higher at the end of the starter and grower periods for 25MD than LD and for both HD and 25MD than LD (P < 0.05). Femur and tibia ash weight were higher for 25MD in comparison to LD birds (P < 0.05) at the end of the grower period. Femur and tibia BBS were higher (P < 0.05) for 25MD in comparison to LD at the end of the grower and finisher periods, respectively. Overall, effects of vitD supply were more pronounced for femur than for tibia mineralization. Results do not suggest supplementation of vitD above current maximum levels and support partial substitution by 25‐OH‐D3.