This study aimed to investigate the effect of eggshell temperature (ET) manipulations during incubation, in ovo feeding (IOF) of arginine, and post-hatch dietary supplementation with guanidinoacetic acid (GAA) on hatching traits and subsequent growth and physiological performance of hatched broiler chicks reared under subnormal temperature. In experiment 1, from d 8 of incubation onward, a total of 2,160 hatching eggs were randomly arranged in a 2 × 3 factorial design, in which the eggs were exposed to 2 ET (37.8°C or periodically low ET), and 3 IOF treatments (noninjected, diluent-injected, and 1% arginine solution-injected). In experiment 2, a total of 576 one-day-old male broiler chicks from 2 temperature conditions and 2 IOF treatment groups (noninjected and Arg-injected) were reared for 42 d with or without GAA supplementation in a 2 × 2 × 2 factorial design. Each treatment had 6 replicates with 12 birds each. A subnormal ambient temperature (17°C) was applied from 15 d onward to induce ascites. Results from experiment 1 showed a 2-way interaction between ET and IOF for embryonic mortality rate during 19 to 21 d of incubation and residual yolk weight at hatch (P < 0.05). A periodically low ET significantly increased yolk free body mass, first-grade chicks, and relative heart weight than an ET of 37.8°C. In the second experiment, overall average daily gain (ADG) was increased, but feed conversion ratio (FCR), ascites mortality, and serum thyroid hormones and corticosterone were reduced in the low ET group (P < 0.05). There were also IOF × GAA interactions for ADG and FCR (P < 0.05). IOF of arginine or dietary GAA increased serum nitric oxide concentration and jejunal villus height, but decreased ascites mortality (P < 0.05). In conclusion, a periodically low ET accompanied by IOF of arginine during incubation and posthatch dietary supplementation with GAA could be a useful strategy for improving the chick quality at hatch and subsequent improvements in post-hatc