Several studies have shown that the transition from egg laying to incubation behavior in birds is associated with changes in plasma levels of prolactin and steroid hormones. However, any effect of the tactile and visual input provided by eggs at initiating these hormonal changes has not been fully investigated in wild birds. A few days before yellow-eyed penguins, Megadyptes antipodes, started egg laying, we placed an artificial egg into their nests or under cages next to their nest. We then investigated the effect of the tactile and/or visual stimulus of such an artificial egg on prolactin secretion, steroid hormone levels (total androgen, estradiol and progesterone), brood patch development, incubation onset and clutch size in these penguins. Prolactin levels rose in females in response to having an artificial egg in the nest, while they declined considerably in males. Total androgen concentrations in males were less than 7% of those of control males and the levels prior to egg placement. Brood patch width increased in both males and females. Additionally, an egg in the nest caused yellow-eyed penguin pairs to attend and sit prone on their nest more frequently. Females that initiated egg laying 1 or 2 days after placement of the artificial egg in the nest, laid a full clutch of two eggs, while most other females that were exposed to an artificial egg in their nest, laid only a single egg. In contrast, the visual stimulus of an artificial egg alone (that was placed under a cage) did not influence hormone levels, brood patch development, incubation behavior or clutch size. The stimulation of an egg in the nest influences prolactin and total androgen levels in yellow-eyed penguins, particularly in males. While brood patch development and incubation behavior were initiated and egg laying was terminated in response to an artificial egg in the nest, the exact endocrine mechanisms underlying these physiological and behavioral changes remain poorly understood. We encourage further studies on other bird species taking an experimental approach to investigate the direct influence of hormones in initiating brood patch development and incubation behavior. Moreover, such experimental studies will widen our understanding of the endocrine mechanisms that regulate clutch size.