How does visible light flicker impact laying hen pullet behavior, fear, and stress levels?

Many characteristics of artificial light have been evaluated; however, light-flicker frequency (F) has not been assessed extensively in poultry. Pullets (1,344 per strain [S]; Lohmann Brown-Lite [LB] and LSL-Lite [LW]) were placed into 8 light-tight rooms, containing 6 floor pens (15 pen replicates per F × S for 30 and 250 Hz; 18 pen replicates per F x S for 90 Hz), and assigned 1 of 3 F treatments (30, 90, 250 Hz). The experiment took place over 2 trials (blocks). To evaluate long-term effects of F during rearing, birds were followed through the hen phase. Data were analyzed using Proc Mixed (SAS 9.4). Differences were considered significant when P ≤ 0.05, and behaviors are expressed as percentage of time. Pullets reared under 30 Hz spent more time performing nutritive behaviors (P < 0.01) and as "unidentified" (P = 0.02) than other treatments. Active behavior demonstrated an age x F interaction, with pullets being more active at 16 wk, regardless of F (P < 0.01). Comfort behaviors were higher at 16 wk compared to other ages, regardless of F (P < 0.01). Exploratory behaviors were lowest at 4 wk in pullets under 30 Hz (P < 0.01). Aggressive behaviors (12 wk) were higher in pullets reared under 250 Hz than those under 90 Hz (P < 0.01). Comb score was unaffected by F (P = 0.79) and all birds scored had a full plumage. Heterophil-to-lymphocyte ratio was unaffected by F at 7 or 15 wk (P = 0.85 and P = 0.54, respectively). In trial 1, pullets reared under 90 Hz had higher corticosterone concentrations than those reared under 250 Hz (P = 0.02) and trial 2 there were no effects of F (P = 0.97). For novel object test, LW pullets reared under 90 Hz had a higher latency to peck than LW pullets under 30 Hz or 250 Hz (P = 0.03). Hen behavior (wk 39) and fear tests (36 wk; novel object test (P = 0.86) and tonic immobility (P = 0.37)) were unaffected by F. Overall, minimal effects of F were seen on pullet and hen behavior and stress.