Variation in advertisement call structure of whistling frogs, Litoria ewingi, was studied among breeding sites in the vicinity of Christchurch on the East coast and Harihari on the West coast of the South Island. Males vocalized in breeding choruses throughout the year and females moved toward and appeared to evaluate calls of territorial males before initiating amplexus. Acoustic interactions between neighbours led to modification of call structure and timing, and wrestling matches between males were frequent. I quantified call structure for 1623 calls of 168 individuals using 24 call parameters. Most temporal characteristics were significantly correlated with the caller's body temperature and dominant frequency was negatively correlated with body size. These results are consistent with the prediction of temperature-dependent metabolic rates in a poikilotherm's nervous system. I compared call structure among the study populations using both multivariate and univariate analyses. Advertisement call structure showed highly significant variation among populations for most call variables. The greatest difference occurred between the West and East coast populations, and a clinal trend occurred in the East coast populations on a microgeographic scale. Variation among East coast populations in some variables was comparable to the scale of variation between coasts. These results suggest that whistling frogs exhibit natal philopatry. I obtained repeated recordings for 21 individuals and examined variation in call structure within and among individuals using nested multivariate analyses of variance. Highly significant variation occurred both within and among individuals for most call characters, but certain call characters were more variable among individuals. This suggests that whistling frog advertisement calls are individually distinctive.