In order to determine the prevalence of Bartonella henselae becteremia in an Australian cat population we examined blood cultures on a group of Sydney cats. Cats referred to the Concord Animal Hospital for euthanasia were selected randomly for blood culture and serum sampling. Blood samples were lysed and centrifuged and then cultured for up to five weeks. Suspicious colonies were identified biochemically as probable B. henselae. Selected isolates were confirmed as B. henselae using the polymerase chain reaction. Of the cats accrued throughout Sydney, 27/77 (35%) were culture positive for B. henselae, of these 24/59 (40%) were feral cats and 3/18 (16%) were domestic. Most cats in the study were younger than one year (mean 9.9 months). Our study demonstrates that bacteremia with B. henselae is common in the metropolitan cat population and suggests that it is particularly prevalent among feral animals. By contrast Cat Scratch Disease (CSD) is a relatively uncommon clinical diagnosis in the Australian population. Explanations for this discrepancy may include poor transmission, low bacterial virulence and underdiagnosis. It is possible that feral animals are a greater potential risk source for this infection than domestic cats.