Spotted hyaenas (Crocuta crocuta) experience a range of influences that affect their demography and are generally regulated by density-dependent mechanisms. Although widely distributed across sub-Saharan Africa surprisingly few spotted hyaena populations have reliable estimates of population sizes, and almost nothing is known about the trends in hyaena numbers. In most areas, threats due to anthropogenic disturbance or diseases are therefore not possible to evaluate. We calibrated a call-up method designed to achieve unbiased counts of African lions (Panthera leo) to estimate spotted hyaena numbers simultaneously. This greatly reduces the time and cost of surveys for these two dominant large carnivores in African savannas. We evaluated the effect of preferred prey biomass, lion density and disease on spotted hyaena population abundance in the Kruger National Park, South Africa, and found evidence only for an effect of prey biomass. Our results suggest that the persistence of spotted hyaena is not threatened in the Kruger National Park.