The population of puku, Kobus vardonii, at Lake Rukwa, one of only two in Tanzania, is poorly known. A multi‐species dry season game count carried out in Rukwa Game Reserve (RGR) in 2004 estimated a population size of 1729, but was associated with a high margin of error [coefficient of variation (CV) 0.88]. To increase the precision for the local puku population estimate, we conducted a zig‐zag line transect survey restricted only to areas occupied by puku in the dry season of 2006. This survey technique produced an estimate of 776 individuals and the CV was considerably reduced to 0.29. Puku herd size was comparable with the studies carried out in the Kilombero valley, Tanzania and in Kasanka National Park, Zambia, thus broadly confirming the herding ecology of this species. We found that pukus were restricted to the central parts of the floodplains, suggesting an avoidance of habitats used by pastoralists at the edge of RGR. Compared with the estimates of the puku population made in the 1980s, the puku population has declined. We recommend regular monitoring and enforcement of the current management plan to conserve the local puku population.