Urban Norway and black rats (Rattus norvegicus and Rattus rattus) are reservoirs for variety of zoonotic pathogens. Many of these pathogens, including Rickettsia typhi, Bartonella spp., and Seoul hantavirus (SEOV), are thought to be endemic in rat populations worldwide; however, past field research has found these organisms to be absent in certain rat populations. Rats (Rattus spp.) from an inner city neighborhood of Vancouver, Canada, were tested for exposure to and/or infection with SEOV and R. typhi (using serology and PCR), as well as Bartonella spp. (using culture and sequencing). Approximately 25% of 404 rats tested were infected with Bartonella tribocorum, which demonstrated significant geographic clustering within the study area. Infection was associated with both season and sexual maturity. Seroreactivity against R. typhi and SEOV was observed in 0.36% and 1.45% of 553 rats tested, respectively, although PCR screening results for these pathogens were negative, suggesting that they are not endemic in the study population. Overall, these results suggest that the geographic distribution of rat-associated zoonoses, including R. typhi, SEOV, and Bartonella spp., is less ubiquitous than previously appreciated, and is likely dependent on patterns of dispersion and establishment of the rat reservoir host. Further study on global and local Rattus spp. population structures may help to elucidate the ecology of zoonotic organisms in these species.