Assessing the physiological stress responses of wild animals opens a window for understanding how organisms cope with environmental challenges. Since stress response is associated with changes in body temperature, the use of body surface temperature through thermal imaging could help to measure acute and chronic stress responses non-invasively. We used thermal imaging, acute handling-stress protocol and an experimental manipulation of glucocorticoid hormone levels in breeding king penguins (Aptenodytes patagonicus), to assess: 1. the potential contribution of the Hypothalamo-Pituitary-Adrenal (HPA) axis in mediating chronic and acute stress-induced changes in adult surface temperature, 2. the influence of HPA axis manipulation on parental investment through thermal imaging of eggs and brooded chicks, and 3. the impact of parental treatment on offspring thermal response to acute handling. Eye temperature (Teye) increased and beak temperature (Tbeak) decreased in response to handling stress in adults, but neither basal nor stress-induced surface temperatures were significantly affected by glucocorticoid implants. While egg temperature was not significantly influenced by parental treatment, we found a surprising pattern for chicks: chicks brooded by the (non-implanted) partner of glucocorticoid-implanted individuals exhibited higher surface temperature (both Teye and Tbeak) than those brooded by glucocorticoid-implanted or control parents. Chick response to handling in terms of surface temperature was characterized by a drop in both Teye and Tbeak independently of parental treatment. We conclude that the HPA seems unlikely to play a major role in determining chronic or acute changes in surface temperature in king penguins. Changes in surface temperature may primarily be mediated by the Sympathetic-Adrenal-Medullary axis (SAM) in response to stressful situations. Our experiment did not reveal a direct impact of parental HPA manipulation on parental investment (egg or chick temperature), but a potential influence on the partner brooding behaviour (i.e. family-transmitted stress hypothesis).