Root-feeding grubs (Coleoptera: Scarabaeidae) were sampled from damaged areas of 61 irrigated roughs on 32 Kentucky golf courses to determine species composition and natural enemy incidence, the first such survey in the United States' transitional turfgrass climatic zone. Masked chafers (Cyclocephala lurida Bland and C. borealis Arrow) and Japanese beetle (Popillia japonica Newman) accounted for ≈73 and 26% of grubs found in an autumn survey, with Cyclocephala spp. predominating at most sites, although mixed infestations were common. Only a few Phyllophaga spp., and no exotic species other than P. japonica were found. Cyclocephala spp. also predominated in seasonal and statewide surveys regardless of whether a course had cool- or warm-season grass fairways. Pathogenic bacteria, Paenibacillus and Serratia spp., and the autumn-active parasitoid Tiphia pygidialis Allen were the main enemies associated with Cyclocephala spp. Predominant enemies of P. japonica were Paenibacillus, Serratia, and Metarhizium spp. in autumn, and eugregarines, Stictospora sp. (probably S. villani Hays and Clopton) and Tiphia vernalis Rohwer in spring. Entomopathogenic nematodes and the microsporidian Ovavesicula popilliae Andreadis & Hanula were nearly absent in our samples. No predictive relationships were found between soil parameters and proportionate abundance of Cyclocephala or P. japonica, or with natural enemy incidence at particular sites. Although incidence of individual enemies was generally low (<20%; often <5%) in these point-in-time surveys, collectively and over their hosts' prolonged development they may take a significant toll on grub populations.