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-hatch performance and ascites indices in cold-stressed broilers.